Carbapenem Antibacterials with Gram-Negative Activity

ABSTRACT

The present invention provides β-methyl carbapenem compounds and pharmaceutical compositions useful in the treatment of bacterial infections and methods for treating such infections using such compounds and/or compositions. The invention includes administering an effective amount of a carbapenem compound or salt and/or prodrug thereof to a host in need of such a treatment.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No.61/356,398, filed Jun. 18, 2010.

FIELD OF THE INVENTION

This application provides novel carbapenem compounds and their salts,methods of treatment of gram-negative bacterial infections with aneffective amount of the compounds and pharmaceutical compositionsincluding the compounds.

BACKGROUND

The worldwide exploitation of antibiotics to treat infectious diseaseshas grown dramatically over the last forty years. In 1954, two millionpounds of antibiotics were produced in the United States. Today, thefigure exceeds 50 million pounds. According to the Centers DiseaseControl (CDC), humans consume 235 million doses of antibiotics annually.

Widespread misuse or overuse of antibiotics has fostered the spread ofantibiotic resistance and has contributed to the development of aserious public health problem. Antibiotic resistance occurs whenbacteria that cause infection are not killed by the antibiotics taken tostop the infection. The bacteria survive and continue to multiply,causing more harm. For example, the bacterium Staphylococous aureus is amajor cause of hospital acquired infections that, historically,responded satisfactorily to the antibiotic vancomycin. Recently,however, many strains of S. aureus have been found to be resistant tovancomycin. Moreover, the death rates for some communicable diseasessuch as tuberculosis have started to rise again, in part because ofincreases in bacterial resistance to antibiotics.

Antibiotics are used therapeutically to treat bacterial infections.Several types of antibiotics, classified according to their mechanism ofaction, are currently employed. The known types of antibiotics include,e.g. cell wall synthesis inhibitors, cell membrane inhibitors, proteinsynthesis inhibitors and inhibitors that bind to or affect the synthesisof DNA or RNA.

Cell wall synthesis inhibitors, such as beta lactam antibiotics,generally inhibit some step in the synthesis of bacterial peptidoglycan.Penicillin is generally effective against non-resistant streptococcus,gonococcus and staphylococcus. Amoxycillin and Ampicillin have broadenedspectra against Gram-negative bacteria. Cephalosporins are generallyused as penicillin substitutes, against Gram-negative bacteria and insurgical prophylaxis. Monobactams are generally useful for the treatmentof allergic individuals.

Numerous antibiotic agents, suitable for use in the treatment ofbacteria-related diseases and disorders, are known and disclosed, e.g.in The Physician's Desk Reference (PDR), Medical Economics Company(Montvale, N.J.), (53^(rd) Ed.), 1999; Mayo Medical Center Formulary,Unabridged Version, Mayo Clinic (Rochester, Minn.), January 1998; MerckIndex: An Encyclopedia of Chemicals, Drugs and Biologicals, (11^(th)Ed.), Merck & Co., Inc. (Rahway, N.J.), 1989; University of WisconsinAntimicrobial Use Guide,http://www.medsch.wisc.edu/clinsci/5amcg/amcg.html; Introduction on theUse of the Antibiotics Guideline, of Specific Antibiotic Classes, ThomasJefferson University,http://jeffiine.tju.edu/CWIS/OAC/antibiotics_guide/intro.html; andreferences cited therein.

The first carbapenem to be isolated was thienamycin, shown below, whichwas isolated from Streptomyces cattleya (U.S. Pat. No. 3,950,357) andwas shown to have strong antibacterial activity, including potencyagainst Pseudomonas spp. and β-lactamase stability (Kahan, J. S., etal., J. Antibiot., 32, pp. 1-12 (1979); Bodey, G. P., et al.,Antimicrob. Agents Chemother., 15, pp. 518-521 (1979). The racernicsynthesis of thienamycin was reported shortly thereafter by Merck(Johnston, D. B. R., et al., J. Am. Chem. Soc., 100, pp. 313-315 (1978);Bouffard, F. A., et al., J. Org. Chem., 45, 1130-1142 (1980)), as wellas an asymmetric total synthesis (Salzmann, T. N., et al., J. Am. Chem.Soc. 102, pp. 6161-6163 (1980)). The nucleus and amino-containing sidechain of this molecule,

however, contributed to its chemical instability. In addition to itspotential to be hydrolyzed by the zinc-activated β-lactamase that ispresent in Bacillus species, Xanthomonas, Pseudomonas, and Bacteroidesspecies (Saino, Y., et al., Antimicrob. Agents Chemother., 22, pp.564-570 (1982); Yotsujii, A., et al., Antimicrob. Agents Chemother., 24,pp. 925-929 (1983)), chemical stability issues associated with theintermolecular aminolysis of the azetidinone (β-lactam) ring of onemolecule of thienamycin by the primary amine in the cysteamine sidechain of another thienamycin molecule, resulted in the use ofthienamycin as a drug candidate to be abandoned.

As a result of the problems associated with thienamycin, N-formimidoylthienamycin, known as imipenem, was synthesized (Leanza, W. J., et al.,J. Med. Chem., 22, pp. 1435-1436 (1979)). This compound bears a morebasic amidine functionality on the 2′ side chain, which is protonated atphysiological pH, preventing the compound from initiating a nucleophilicattack on another imipenem molecule.

However, poor urinary tract recovery from test subjects revealed aninstability of this compound to the mammalian β-lactamase renaldehydropeptidase-I (DHP-I) (Shimada, J., et al., Drugs Exp Clin Res.,20, pp. 241-245 (1994)). Consequently, the compound cilastatin wasdeveloped for use in co-administration in order to prevent hydrolysisand degredation by DHP-I; this combination therapy is currentlyprescribed under the name Primaxin® (Merck Frosst Std).

In response to the problem of carbapenems to destruction by renaldehydropeptidase-1, the carbapenem antibiotic meropenem (SM7338) (shownbelow), was developed (see, Edwards, J. R., et al., Antimicrob. AgentsChemother., 33, pp. 215-222 (1989); Neu, H. C., et al., Antimicrob.Agents Chemother., 33, pp. 1009-1018 (1989)).

This compound was shown to be active against a large number ofGram-negative bacteria. The drug is currently prescribed for intravenoususe (Merrem® IV; AstraZeneca) in the treatment of intra-abdominalinfections and bacterial meningitis.

The carbapenem ertapenem (formerly MK-0826; Cunha, B. A., Drugs ofToday, 38, pp. 195-213 (2002)) was the first of a group of carbapenemswith potential against methicillin-resistant staphylococci (MRS) shownto be useful as a long-acting, parenteral carbapenem (Shah, P. M., etal., J. Antimicrob. Chemother., 52, pp. 538-542 (2003); Aldridge, K. E.,Diagn. Microbiol. Infect. Dis., 44(2), pp. 181-6 (2002)). It is suitablefor administration both as a single-agent (e.g., co-administration witha compound such as cilastatin is not required), or by the intravenous orintramuscular route (Legua, P., et al., Clin. Therapeut., 24, pp.434-444 (2002); Majumdar, A. K., et al., Antimicrob. Agents Chemother.,46, pp. 3506-3511 (2002)). Ertapenem has received regulatory approval inboth the United States (November, 2001) and the European Union (April,2002).

One carbapenem having a fused pyrazole ring system (L-627; Biapenem) wasdeveloped by Lederle Ltd. (Japan), and introduced a methyl radical atthe 1-β postion of the carbapenem skeleton (see, U.S. Pat. No.4,866,171). This structural modification reportedly gave biapenemstability against hydrolysis by kidney dehydropeptidase, makingco-administration of a dehydropeptidase inhibitor unnecessary.

More recently, a new, injectable 1-β-methyl carbapenem antibiotic havingan (R)-1-hydroxymethyl-methylaminopropyl group exhibiting both broadspectrum, potent antibacterial activity (BO-2727) and havingantipseudomonal activity has been reported (Nakagawa, S., et al.,Antimicrob. Agents Chemother., 37, pp. 2756-2759 (1993); Hazumi, N., etal., Antimicrob. Agents Chemother., 39, pp. 702-706 (1995).

Since the discovery of thienamycin having a potential antimicrobialactivity against Gram-negative and Gram-positive bacteria, studies onthe syntheses of carbapenem derivatives which are analogous tothienamycin have been widely developed. As a result, it was found thatcarbapenem derivatives having, as their 2-side chain, a substituentderived from 4-hydroxy-proline exhibit a potential antimicrobialactivity and are useful as medicines or as intermediates for compoundspossessing antimicrobial activity.

1-β-methyl carbapenem antibiotics, are particularly well known fortreating a broad spectrum of gram-negative and gram-positive bacterialinfections. See for example U.S. Pat. No. 4,962,103; U.S. Pat. No.4,933,333; U.S. Pat. No. 4,943,569; U.S. Pat. No. 5,122,604; U.S. Pat.No. 5,034,384 and U.S. Pat. No. 5,011,832.

U.S. Pat. No. 6,255,300 to Merck & Co. describes certain carbapenemantibacterial agents in which the carbapenem nucleus is substituted withan iodo-phenyl linked through a methyl-oxygen linkage. The patent statesthat these compounds are useful against gram positive bacterialinfections. Similarly, U.S. Pat. No. 6,310,055 provides carbapenemcompounds with aromatic side chains that are halogen substituted, linkedthorough an alkoxy unsaturated group.

European Publication No. 0 292 191 to Merck & Co. describes certain2-(substituted methyl)-1-alkylcarbapenem compounds useful as antibioticagents.

U.S. Pat. No. 6,399,597, also to Merck & Co. describes certainnapthosultam compounds that are allegedly useful in the treatment ofcertain drug resistant bacterial infections.

U.S. Pat. No. 7,683,049 to FOB Synthesis, Inc. describes certainβ-methyl carbapenem compounds for the treatment of gram-negativebacterial infections.

Because of the drug-resistance challenges associated with treatingbacterial infections, there remains a need for new antimicrobial agents.

Therefore, it is one object of the present invention to provide novel3-methylcompounds carbapenems that are effective antimicrobial agents.

It is another object of the present invention to provide methods for thetreatment of gram-negative bacteria, which optionally can bedrug-resistant and/or multi-drug resistant.

SUMMARY OF THE INVENTION

In one embodiment of the present invention, carbapenem compounds of thegeneral

or a pharmaceutically acceptable salt, ester or prodrug thereof, aredescribed,whereinR¹ and R² are each independently selected from H or alkyl;P is H, OH, halogen, or hydroxyl protected by a hydroxyl protectinggroup;n is 0, 1 or 2;X is —(CR₂)_(m)— or —C(═O)—;m is 0, 1 or 2;

Y is CN, OR, SR′ or NRR′;

each R is independently selected from H, alkyl or haloalkyl; andR′ is H, alkyl, NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂;CR₂C(═O)NR₂; C(═NR)R; C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; orC(═O)CR₂NRC(═NR)NR₂; andZ is H, alkyl, halo, CN, OR, SR′ or NRR′.

In another embodiment, carbapenem compounds of Formula IV:

or a pharmaceutically acceptable salt, ester or prodrug thereof, aredescribedwhereinR¹, R² and R³ are each independently selected from H or alkyl;P is H, OH, halogen, or hydroxyl protected by a hydroxyl protectinggroup;X is —(CR₂)_(m)— or —C(═O)—;m is 0, 1 or 2;

Y is CN, OR, SR or NRR′;

each R is independently selected from H, alkyl or haloalkyl; andR′ is H, alkyl, NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂;CR₂C(═O)NR₂; C(═NR)R; C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; orC(═O)CR₂NRC(═NR)NR₂; andZ is H, alkyl, halo, CN, OR, SR′ or NRR′.

In another embodiment, carbapenem compounds of Formula VI:

or a pharmaceutically acceptable salt, ester or prodrug thereof, aredescribedwhereinR¹ and R² are each independently selected from H or alkyl;P is H, OH, halogen, or hydroxyl protected by a hydroxyl protectinggroup;X is —(CR₂)_(m)— or —C(═O)—;m is 0, 1 or 2;

Y is CN, OR, SR′ or NRR′;

each R is independently selected from H, alkyl or haloalkyl;R′ is H, alkyl, NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂;CR₂C(═O)NR₂; C(═NR)R; C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; orC(═O)CR₂NRC(═NR)NR₂; andZ is H, alkyl, halo, CN, OR, SR′ or NRR′.

In a particular embodiment, the present invention describes thefollowing compound:

In another particular embodiment, the present invention describes thefollowing compound:

The present invention also provides a pharmaceutical compositionincluding a compound of the invention, or a pharmaceutically acceptablesalt and/or prodrug thereof, optionally with a pharmaceuticallyacceptable carrier or diluent.

In one embodiment, the invention provides a pharmaceutical compositioncomprising a compound of the invention, or a pharmaceutically acceptablesalt and/or prodrug therein, in combination with one or more otherantimicrobial agents, optionally with a pharmaceutically acceptablecarrier or diluent.

In another embodiment, the invention provides a method of preventing ortreating a bacterial infection in a host, typically an animal, and mosttypically a human, including administering to the host a therapeuticamount of a compound of the present invention, or a pharmaceuticallyacceptable salt and/or prodrug therein, optionally in a pharmaceuticallyacceptable carrier or diluent.

In a separate embodiment, the invention provides a method of preventingor treating a gram-negative bacterial infection in a host that includesadministering a therapeutic amount of a compound of the presentinvention, or a pharmaceutically acceptable salt and/or prodrug therein,in combination or alternation with one or more other antimicrobialagents, optionally in a pharmaceutically acceptable carrier or diluent.

In one principal embodiment, the bacterial infection is due to agram-negative bacteria. In another embodiment, the bacterial infectionis from a drug resistant and/or multiple-drug resistant gram-negativebacteria.

The invention also provides a compound of the present invention for usein medical therapy, and the use in the preparation of a medicament forthe treatment of bacterial infections, particularly gram negativebacterial infections, alone or in combination with another agent.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides carbapenem compounds or their pharmaceuticallyacceptable salts or prodrugs, pharmaceutical compositions containingthese compounds and methods of their use in the treatment or preventionof gram-negative bacterial infections.

DEFINITIONS

The numbering system for the carbapenem compounds used in thisspecification is set out below, wherein the numbering of the carbapenemnucleus is in accordance with standards in the art (see, Tiraby, G., etal., Biochem J, 276 (pt. 1), pp. 269-270 (1991)).

Whenever a range is presented herein it should be understood to includeeach element of the range. For example, the range “C₁ to C₄” alkylindependently includes C₁, C₂, C₃ and C₄ alkyl groups. When such a rangeis stated, each element has been contemplated and the range is usedmerely for convenience.

Generally, while the compounds, compositions and methods are describedin terms of “comprising” various components or steps, the compounds,compositions and methods can also “consist essentially of” or “consistof” the various components and steps.

The term “alkyl”, as used herein, unless otherwise specified, includes asaturated straight, branched, or cyclic, primary, secondary, or tertiaryhydrocarbon of C₁ to C₁₀. The term includes both substituted andunsubstituted alkyl groups. Moieties with which the alkyl group can besubstituted are selected from the group consisting of hydroxyl, halo (F,Cl, Br, I), amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano,sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate,either unprotected, or protected as necessary, as known to those skilledin the art, for example, as taught in Greene, et al., Protective Groupsin Organic Synthesis, John Wiley and Sons, Second Edition, 1991, herebyincorporated by reference. When the alkyl group is said to besubstituted with an alkyl group, this is used interchangeably with“branched alkyl group”. Specific examples of alkyls and/or substitutedalkyls includes, but are not limited to, methyl, trifluoromethyl, ethyl,propyl, isopropyl, cyclopropyl, butyl, isobutyl, t-butyl, pentyl,cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl,cyclohexylmethyl, 3-methylpentyl, 2,2-dimethylbutyl, and2,3-dimethylbutyl.

The term “lower alkyl”, as used herein, and unless otherwise specified,refers to a C₁ to C₄ saturated straight, branched, or if appropriate, acyclic (for example, cyclopropyl) alkyl group, including bothsubstituted and unsubstituted forms. Unless otherwise specificallystated in this application, when alkyl is a suitable moiety, lower alkylis typical. Similarly, when alkyl or lower alkyl is a suitable moiety,unsubstituted alkyl or lower alkyl is typical.

Cycloalkyl is a species of alkyl containing from 3 to 15 carbon atoms,without alternating or resonating double bonds between carbon atoms. Itmay contain from 1 to 4 rings, which are fused.

The term “alkenyl” includes a hydrocarbon radical straight, branched orcyclic containing from 2 to 10 carbon atoms and at least one carbon tocarbon double bond. Examples of alkenyl groups include ethenyl,propenyl, butenyl and cyclohexenyl.

The term “alkynyl” refers to a hydrocarbon radical straight or branched,containing from 2 to 10 carbon atoms and at least one carbon to carbontriple bond. Examples of alkynyl groups include ethynyl, propynyl andbutynyl.

“Alkoxy” includes C₁-C₄ alkyl-O—, with the alkyl group optionallysubstituted as described herein.

The term “alkylamino” or “arylamino” refers to an amino group that hasone or two alkyl or aryl substituents, respectively.

“Aryl” refers to aromatic rings e.g., phenyl, substituted phenyl,biphenyl, and the like, as well as rings which are fused, e.g.,naphthyl, phenanthrenyl and the like. An aryl group thus contains atleast one ring having at least 6 atoms, with up to five such rings beingpresent, containing up to 22 atoms therein, with alternating(resonating) double bonds between adjacent carbon atoms or suitableheteroatoms. The typical aryl groups are phenyl, naphthyl andphenanthrenyl. The term includes both substituted and unsubstitutedmoieties. The aryl group can be substituted with one or more moietiesselected from the group consisting of bromo, chloro, fluoro, iodo,hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano,sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate,either unprotected, or protected as necessary, as known to those skilledin the art, for example, as taught in Greene, et al., Protective Groupsin Organic Synthesis, John Wiley and Sons, Second Edition, 1991. Typicalsubstituted aryls include phenyl and naphthyl.

The term “alkaryl” or “alkylaryl” refers to an alkyl group with an arylsubstituent. The term “aralkyl” or “arylalkyl” refers to an aryl groupwith an alkyl substituent.

The term “heteroaryl” or “heteroaromatic”, as used herein, refers to anaromatic group that includes at least one sulfur, oxygen, nitrogen orphosphorus in the aromatic ring. Heteroaryl or heteroaromatic compoundsinclude monocyclic aromatic hydrocarbon group having 5 or 6 ring atoms,or a bicyclic aromatic group having 8 to 10 atoms, containing at leastone heteroatom, O, S or N, in which a carbon or nitrogen atom is thepoint of attachment, and in which one, two or three additional carbonatoms are optionally replaced by a heteroatom selected from oxygen,sulfur or nitrogen heteroatom. Examples of this type are pyrrole,pyridine, oxazole, thiazole and oxazine. Additional nitrogen atoms maybe present together with the first nitrogen and oxygen or sulfur,giving, e.g., thiadiazole. Examples include the following.

The heteroaryl or heteroaromatic group can be optionally substitutedwith one or more substituent selected from halogen, haloalkyl, alkyl,alkoxy, hydroxy, carboxyl derivatives, amido, amino, alkylamino,dialkylamino. Functional oxygen and nitrogen groups on the heterocyclicor heteroaryl group can be protected as necessary or desired. Suitableprotecting groups are well known to those skilled in the art, andinclude trimethylsilyl, dimethylhexylsilyl, t-butyldimethylsilyl, andt-butyl-diphenylsilyl, trityl or substituted trityl, alkyl groups, acylgroups such as acetyl and propionyl, methanesulfonyl, andp-toluenylsulfonyl.

“Heteroarylium” refers to heteroaryl groups bearing a quaternarynitrogen atom and thus a positive charge. Examples include thefollowing.

When a charge is shown on a particular nitrogen atom in a ring, whichcontains one or more additional nitrogen atoms, it is understood thatthe charge may reside on a different nitrogen atom in the ring by virtueof charge resonance that occurs.

The term “heterocycloalkyl” refers to a cycloalkyl group (nonaromatic)in which one of the carbon atoms in the ring is replaced by a heteroatomselected from O, S or N, and in which up to three additional carbonatoms may be replaced by heteroatoms.

The terms “quaternary nitrogen” and “positive charge” refer totetravalent, positively charged nitrogen atoms including, e.g., thepositively charged nitrogen in a tetraalkylammonium group (e. g.tetramethylammonium), heteroarylium, (e.g., N-methyl-pyridinium), basicnitrogens which are protonated at physiological pH, and the like.Cationic groups thus encompass positively charged nitrogen-containinggroups, as well as basic nitrogens which are protonated at physiologicpH.

The term “heteroatom” refers to oxygen, sulfur, nitrogen, phosphorus,and selenium, selected on an independent basis.

Halogen and “halo”, as used herein, includes bromine, chlorine, fluorineand iodine.

The term acyl refers to a carboxylic acid ester in which thenon-carbonyl moiety of the ester group is selected from straight,branched, or cyclic alkyl or lower alkyl, alkoxyalkyl includingmethoxymethyl, aralkyl including benzyl, aryloxyalkyl such asphenoxymethyl, aryl including phenyl optionally substituted withhalogen, C₁ to C₄ alkyl or C₁ to C₄ alkoxy, sulfonate esters such asalkyl or aralkyl sulphonyl including methanesulfonyl, the mono, di ortriphosphate ester, trityl or monomethoxytrityl, substituted benzyl,trialkylsilyl (e.g. dimethyl-t-butylsilyl) or diphenylmethylsilyl. Arylgroups in the esters typically include a phenyl group. The term “loweracyl” refers to an acyl group in which the non-carbonyl moiety is loweralkyl.

“Carboxylate anion” refers to a negatively charged group —COO.

“Guanidinyl” refers to the group: H₂NC(NH)NH—.

“Carbamimidoyl” refers to the group: H₂NC(NH)—.

“Ureido” refers to the group: H₂NC(O)NH—.

When a group is “optionally interrupted”, this includes one or more ofthe interrupting moieties in combination, as well as said moietieslocated at either or both ends of the chain. Thus, it includesterminating the group as well.

When a group is termed “substituted”, unless otherwise indicated, thismeans that the group contains from 1 to 4 substituents thereon. Withrespect to R, R^(a), R^(b) and R^(c), the substituents available onalkyl groups are selected from the values of R^(d). Many of the variablegroups are optionally substituted with up to four R^(i) groups. Withrespect to R^(e), R^(f) and R^(g), when these variables representsubstituted alkyl, the substituents available thereon are selected fromthe values of R^(i).

When a functional group is termed “protected”, this means that the groupis in modified form to preclude undesired side reactions at theprotected site, and unless otherwise defined refers to a group that isadded to an oxygen, nitrogen, or phosphorus atom to prevent its furtherreaction or for other purposes. In some of the carbapenem compounds ofthe present invention, M is a readily removable carboxyl protectinggroup, and/or P represents a hydroxyl which is protected by ahydroxylprotecting group. Such protecting groups are used toprotectively block the hydroxyl or carboxyl group during the synthesisprocedures and are readily removable by procedures that will not causecleavage or other disruption of the remaining portions of the molecule.Such procedures include chemical and enzymatic hydrolysis, treatmentwith chemical reducing or oxidizing agents under mild conditions,treatment with a transition metal catalyst and a nucleophile andcatalytic hydrogenation.

A wide variety of oxygen and nitrogen protecting groups are known tothose skilled in the art of organic synthesis. Suitable protectinggroups for the compounds of the present invention will be recognizedfrom the present application taking into account the level of skill inthe art, and with reference to standard textbooks, such as Greene, T. W.and Wuts, P. M., Protective Groups in Organic Synthesis, 3^(rd) Ed.,Wiley, New York (1991). Examples of carboxyl protecting groups includeallyl, benzhydryl, 2-naphthylmethyl, benzyl (Bn), silyl such ast-butyldimethylsilyl (TBDMS), phenacyl, p-methoxybenzyl, o-nitrobenzyl,p-methoxyphenyl, p-nitrobenzyl, 4-pyridylmethyl and t-butyl. Examples ofsuitable C-6 hydroxyethyl protecting groups include triethylsilyl (TES),t-butyldimethylsilyl (TBDMS), o-nitrobenzyloxycarbonyl (ONB),p-nitrobenzyloxycarbonyl (PNB), benzyloxycarbonyl (CBz),allyloxycarbonyl (Alloc), t-butyloxycarbonyl (Boc),2,2,2-trichloroethyloxycarbonyl (Troc), and the like.

The phrase “pharmaceutically acceptable ester, salt or hydrate,” refersto those salts, esters and hydrated forms of the compounds of thepresent invention, which would be apparent to the pharmaceuticalchemist. i.e., those which are substantially non-toxic and which mayfavorably affect the pharmacokinetic properties of said compounds, suchas palatability, absorption, distribution, metabolism and excretion.Other factors that are also important in the selection are cost of theraw materials, ease of crystallization, yield, stability, solubility,hygroscopicity and flowability of the resulting bulk drug.

“Pharmaceutically acceptable salts” include salts that retain thedesired biological activity of the parent compound and do not impartundesired toxicological effects. These salts can take the form —COOM,where M is a positive charge, which is balanced by a counterion. Theseinclude salts formed with cations such as sodium, potassium, NH₄ ⁺,magnesium, zinc, ammonium, or alkylammonium cations such astetramethylammonium, tetrabutylammonium, choline, triethylhydroammonium,meglumine, triethanolhydroammonium, calcium, and calcium polyamines suchas spermine and spermidine. These can also include salts formed fromelemental anions such as chloride, bromide, and iodide. They can alsoinclude acid addition salts, for example, salts derived from inorganicor organic acids. Included among such salts are the following: acetate,adipate, alginate, ascorbic acid, aspartate, benzoate, benzenesulfonate,bisulfate, butyrate, citrate, camphorate, camphorsulfonate,cyclopentancpropionate, digluconate, dodecylsulfate, ethanesulfonate,fumarate, glucoheptanoate, gluconic acid, glycerophosphate, hemisulfate,heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide,2-hvdroxvethanesulfonate, lactate, maleate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitric acid, oxalate, palmitic acid,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphoric acid,picrate, pivalate, polygalacturonic acid; polyglutamic acid, propionate,p-toluenesulfonic acid, succinate, sulfuric acid, tannic acid, tartrate,thiocyanate, tosylate and undecanoate.

The term “prodrug” includes a compound that, when administered to ananimal, is converted under physiological conditions to a compound of theinvention, for example a pharmaceutically acceptable ester.

The pharmaceutically acceptable esters are such as would be readilyapparent to a medicinal chemist, and include, for example, thosedescribed in detail in U.S. Pat. No. 4,309,438. Included within suchpharmaceutically acceptable esters are those, which are hydrolyzed underphysiological conditions, such as pivaloyloxymethyl, acetoxymethyl,phthalidyl, indanyl and methoxymethyl. These are also referred to as“biolabile esters”, which are biologically hydrolysable. Examples ofbiolabile esters include compounds in which M represents an alkoxyalkyl,alkylcarbonyloxyalkyl, alkoxycarbonyloxyalkyl, cycloalkoxyalkyl,alkenyloxyalkyl, aryloxyalkyl, alkoxyaryl, alkylthioalkyl,cycloalkylthioalkyl, alkenylthioalkyl, arylthioalkyl or alkylthioarylgroup. These groups can be substituted in the alkyl or aryl portionsthereof with acyl or halo groups. The following M species are examplesof biolabile ester forming moieties: acetoxymethyl, 1-acetoxyethyl,1-acetoxypropyl, pivaloyloxymethyl, 1isopropyloxycarbonyloxyethyl,1-cyclohexyloxycarbonyloxyethyl, phthalidyl and (2-oxomethyl-1,3-dioxolenyl)methyl.

The term “host”, as used herein, refers to a unicellular ormulticellular organism in which the bacteria can replicate, includingcell lines and animals. Alternatively, the host can be carrying a partof the bacterial particles, whose replication and/or function can bealtered by the compounds of the present invention. The term host refersto infected cells, cells transfected with all or part of the bacteriaand animals, such as, primates (including chimpanzees) and, in oneembodiment, the host is a human. Veterinary applications are alsoencompassed by the present invention.

The term “treatment” as used herein, includes an approach for obtainingbeneficial or desired results including clinical results, includingalleviation of symptoms, diminishment of extent of disease,stabilization (i.e., not worsening) state of disease, preventing spreadof disease, preventing or reducing occurrence or recurrence of disease,delay or slowing of disease progression, and reduction of incidence ofdisease or symptoms. As used herein, the phrase “anti-bacteriallyeffective amount” means an amount effective for treating the bacterialinfection.

Compounds of the Invention

In one embodiment, the compound is a compound of Formula I,

or a pharmaceutically acceptable salt, ester or prodrug thereof,whereinR¹ and R² are each independently selected from H or alkyl;P is H, OH, halogen, or hydroxyl protected by a hydroxyl protectinggroup;n is 0, 1 or 2;X is —(CR₂)_(m)— or —C(═O)—;m is 0, 1 or 2;

Y is CN, OR, SR′ or NRR′;

each R is independently selected from H, alkyl or haloalkyl;R′ is H, alkyl, NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂;CR₂C(═O)NR₂; C(═NR)R; C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; orC(═O)CR₂NRC(═NR)NR₂; andZ is H, alkyl, halo, CN, OR, SR′ or NRR′.

In one embodiment, R¹ is H. In one embodiment, R¹ is alkyl, for exampleCH₃. In one embodiment, R² is H. In one embodiment, R² is alkyl, forexample CH₃. In one embodiment, both R¹ and R² are alkyl, for exampleCH₃.

In one embodiment, P is H. In one embodiment, P is OH. In oneembodiment, P is halogen. In one embodiment, P is hydroxyl protected bya hydroxyl protecting group. In a particular embodiment, P is OH orhydroxyl protected by a hydroxyl protecting group.

In one embodiment, n is 0. In one embodiment, n is 1. In anotherembodiment, n is 2. In one embodiment, n is 1 or 2. In one embodiment, nis not 0.

In one embodiment, X is —(CR₂)_(m)—. In one subembodiment, m is 0. Inanother subembodiment, m is 1. In another subembodiment, m is 2. In oneembodiment, X is —C(═O)—.

In one embodiment, at least one R is H. In one embodiment, at least twoRs are H. In one embodiment, at least one R is alkyl, for example CH₃,CH₂CH₃ or CH₂CH₂CH₃. In one embodiment, at least one R is haloalkyl, forexample CF₃.

In one embodiment, Y is CN. In another embodiment, Y is OR. In aparticular embodiment, Y is OH. In one embodiment, Y is SR′, for exampleSH or S(alkyl). In one embodiment, Y is SR′ and R′ is C(═NR)NR₂, forexample C(═NH)NH₂, C(═NCH₃)NH₂, C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃)₂,C(═NR)NH(CH₃), or C(═NCF₃)NH₂. In one embodiment, Y is NRR′. In asubembodiment, Y is NHR′. In another subembodiment, Y is N(alkyl)R′, forexample N(CH₃)R′.

In one embodiment, R′ is H. In one embodiment, R′ is alkyl, for exampleCH₃. In one embodiment, R′ is NR₂, for example NH₂, NHR, NHCH₃, orN(CH₃)₂. In another embodiment, R′ is C(═O)R, for example C(═O)CH₃ orC(═O)CF₃. In another embodiment, R′ is SO₂R, for example SO₂CH₃. Inanother embodiment, R′ is SO₂NR₂, for example SO₂NH₂. In anotherembodiment, R′ is C(═NR)NR₂, for example C(═NH)NH₂, C(═NCH₃)NH₂,C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃)₂, C(═NR)NH(CH₃), or C(═NCF₃)NH₂. Inanother embodiment, R′ is C(═O)NR₂, for example C(═O)NH₂, C(═O)NHR,C(═O)NHCH₃ or C(═O)N(CH₃)₂. In another embodiment, R′ is C(═NR)R, forexample C(═NH)H, C(═NH)R or C(═NH)CH₃. In another embodiment, R′ isC(═NR)NRSO₂R, for example C(═NH)NHSO₂H, C(═NH)NHSO₂R, or C(═NH)NHSO₂CH₃.In another embodiment, R′ is C(═NR)NRC(═O)R, for example C(═NH)NHC(═O)Ror C(═NH)NHC(═O)CH₃. In another embodiment, R′ is C(═O)CR₂NRSO₂NR₂, forexample C(═O)CH₂NHSO₂NR₂, C(═O)CH₂NHSO₂NH₂, or C(═O)CH₂NHSO₂N(CH₃)₂. Inanother embodiment, R′ is C(═O)CR₂NRC(═NR)NR₂, for exampleC(═O)CH₂NHC(═NH)NR₂, C(═O)CH₂NHC(═NH)NH₂ or C(═O)CH₂NHC(═NH)N(CH₃)₂.

In certain embodiments, Y is CN, NR₂, SC(═NR)NR₂, C(═O)NR₂; C(═O)NRSO₂R;C(═O)NRSO₂NR₂; NRC(═NR)NR₂; NRSO₂NR₂; NRC(O)NR₂; NRCR₂C(O)NR₂;NRCR(═NR); CR₂NRC(═NR)NR₂; NRC(═NR)NRSO₂R; NRC(═NR)NRC(O)R;C(O)NRCR₂C(O)NR₂; C(O)NRC(═NR)NR₂; OR; NRC(O)CR₂NRSO₂NR₂; orNRC(O)CR₂NRC(═NR)NR₂.

In one embodiment, Z is H. In another embodiment, Z is alkyl. In oneembodiment, Z is CN. In another embodiment, Z is halo. In certainembodiments, Z is OR, for example OH.

In one embodiment, Z is SR′, for example SH or S(alkyl). In oneembodiment, Z is NRR′. In a subembodiment, Z is NHR′. In anothersubembodiment, Z is N(alkyl)R′, for example N(CH₃)R′.

In one embodiment, R¹ is alkyl; R² is alkyl; P is hydroxyl or hydroxylprotected by a hydroxyl protecting group; n is 0, 1 or 2; m is 0; and Yis —CN. In another embodiment, R¹ is alkyl; R² is alkyl; P is hydroxylor hydroxyl protected by a hydroxyl protecting group; n is 0, 1 or 2; mis 0; and Y is OR. In another embodiment, R¹ is alkyl; R² is alkyl; P ishydroxyl or hydroxyl protected by a hydroxyl protecting group; n is 0, 1or 2; m is 0 or 1; and Y is NRR′.

In one embodiment, when Y is CN, X is not —C(═O)—. In anotherembodiment, R¹ is alkyl; R² is alkyl; P is hydroxyl or hydroxylprotected by a hydroxyl protecting group; n is 0, 1 or 2; X is —C(═O)—;and Y is OR. In another embodiment, R¹ is alkyl; R² is alkyl; P ishydroxyl or hydroxyl protected by a hydroxyl protecting group; n is 0, 1or 2; X is —C(═O)—; and Y is NRR′.

In one embodiment, R¹ is alkyl; R² is alkyl; P is hydroxyl or hydroxylprotected by a hydroxyl protecting group; n is 1 or 2; m is 0; and Y is—CN. In another embodiment, R¹ is alkyl; R² is alkyl; P is hydroxyl orhydroxyl protected by a hydroxyl protecting group; n is 1 or 2; m is 0;and Y is OR. In another embodiment, R¹ is alkyl; R² is alkyl; P ishydroxyl or hydroxyl protected by a hydroxyl protecting group; n is 1 or2; m is 0 or 1; and Y is NRR′. In one embodiment, R¹ is alkyl; R² isalkyl; P is hydroxyl or hydroxyl protected by a hydroxyl protectinggroup; n is 1 or 2; X is —C(═O)—; and Y is OR. In another embodiment, R¹is alkyl; R² is alkyl; P is hydroxyl or hydroxyl protected by a hydroxylprotecting group; n is 1 or 2; X is —C(═O)—; and Y is NRR′.

In one embodiment, the compound of Formula I is selected from the groupconsisting of:

In one embodiment, the compound is compound 7. In another embodiment,the compound is compound 12. In another embodiment, the compound iscompound 19. In another embodiment, the compound is compound 27. Inanother embodiment, the compound is compound 32. In another embodiment,the compound is compound 43. In another embodiment, the compound iscompound 46. In another embodiment, the compound is compound 49. Inanother embodiment, the compound is compound 64. In another embodiment,the compound is compound 89. In another embodiment, the compound iscompound 99. In another embodiment, the compound is compound 137. Inanother embodiment, the compound is compound 146. In another embodiment,the compound is compound 151. In another embodiment, the compound iscompound 156.

In another embodiment, the compound is a compound of Formula II,

or a pharmaceutically acceptable salt, ester or prodrug thereof,whereinR¹ and R² are each independently selected from H or alkyl;P is H, OH, halogen, or hydroxyl protected by a hydroxyl protectinggroup;X is —(CR₂)_(m)— or —C(═O)—;m is 0, 1 or 2;

Y is CN, OR, SR′ or NRR′;

each R is independently selected from H, alkyl or haloalkyl;R′ is H, alkyl, NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂;CR₂C(═O)NR₂; C(═NR)R; C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; orC(═O)CR₂NRC(═NR)NR₂; andZ is H, alkyl, halo, CN, OR, SR′ or NRR′

In one embodiment, R¹ is H. In one embodiment, R¹ is alkyl, for exampleCH₃. In one embodiment, R² is H. In one embodiment, R² is alkyl, forexample CH₃. In one embodiment, both R¹ and R² are alkyl, for exampleCH₃.

In one embodiment, P is H. In one embodiment, P is OH. In oneembodiment, P is halogen. In one embodiment, P is hydroxyl protected bya hydroxyl protecting group. In a particular embodiment, P is OH orhydroxyl protected by a hydroxyl protecting group.

In one embodiment, X is —(CR₂)_(m)—. In one subembodiment, m is 0. Inanother subembodiment, m is 1. In another subembodiment, m is 2. In oneembodiment, X is —C(═O)—.

In one embodiment, at least one R is H. In one embodiment, at least twoRs are H. In one embodiment, at least one R is alkyl, for example CH₃,CH₂CH₃ or CH₂CH₂CH₃. In one embodiment, at least one R is haloalkyl, forexample CF₃.

In one embodiment, Z is H. In one embodiment, Z is halogen. In anotherembodiment, Z is alkyl. In one embodiment, Z is CN. In anotherembodiment, Z is halo. In certain embodiments, Z is OR, for example OH.In one embodiment, Z is SR′, for example SH or S(alkyl). In oneembodiment, Z is halogen, for example mono- or multi-F or Cl. In oneembodiment, Z is NRR′. In a subembodiment, Z is NHR′. In anothersubembodiment, Z is N(alkyl)R′, for example N(CH₃)R′.

In one embodiment, Y is CN. In another embodiment, Y is OR. In aparticular embodiment, Y is OH. In one embodiment, Y is SR′, for exampleSH or S(alkyl). In one embodiment, Y is SR′ and R′ is C(═NR)NR₂, forexample C(═NH)NH₂, C(═NCH₃)NH₂, C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃)₂,C(═NR)NH(CH₃), or C(═NCF₃)NH₂. In one embodiment, Y is NRR′. In asubembodiment, Y is NHR′. In another subembodiment, Y is N(alkyl)R′, forexample N(CH₃)R′.

In one embodiment, R′ is H. In one embodiment, R′ is alkyl, for exampleCH₃. In one embodiment, R′ is NR₂, for example NH₂, NHR, NHCH₃, orN(CH₃)₂. In another embodiment, R′ is C(═O)R, for example C(═O)CH₃ orC(═O)CF₃. In another embodiment, R′ is SO₂R, for example SO₂CH₃. Inanother embodiment, R′ is SO₂NR₂, for example SO₂NH₂. In anotherembodiment, R′ is C(═NR)NR₂, for example C(═NH)NH₂, C(═NCH₃)NH₂,C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃) 2, C(═NR)NH(CH₃), or C(═NCF₃)NH₂. Inanother embodiment, R′ is C(═O)NR₂, for example C(═O)NH₂, C(═O)NHR,C(═O)NHCH₃ or C(═O)N(CH₃)₂. In another embodiment, R′ is C(═NR)R, forexample C(═NH)H, C(═NH)R or C(═NH)CH₃. In another embodiment, R′ isC(═NR)NRSO₂R, for example C(═NH)NHSO₂H, C(═NH)NHSO₂R, or C(═NH)NHSO₂CH₃.In another embodiment, R′ is C(═NR)NRC(═O)R, for example C(═NH)NHC(═O)Ror C(═NH)NHC(═O)CH₃. In another embodiment, R′ is C(═O)CR₂NRSO₂NR₂, forexample C(═O)CH₂NHSO₂NR₂, C(═O)CH₂NHSO₂NH₂, or C(═O)CH₂NHSO₂N(CH₃)₂. Inanother embodiment, R′ is C(═O)CR₂NRC(═NR)NR₂, for exampleC(═O)CH₂NHC(═NH)NR₂, C(═O)CH₂NHC(═NH)NH₂ or C(═O)CH₂NHC(═NH)N(CH₃)₂.

In a particular embodiment, Y is NH₂, NHC(═NH)NH₂, or NHSO₂NH₂.

In one embodiment, the compound of Formula II is selected from the groupconsisting of

In another embodiment, the compound is a compound of Formula III,

or a pharmaceutically acceptable salt, ester or prodrug thereof,whereinR¹ and R² are each independently selected from H or alkyl;P is H, OH, halogen, or hydroxyl protected by a hydroxyl protectinggroup;X is —(CR₂)_(m)— or —C(═O)—;m is 0, 1 or 2;

Y is CN, OR, SR′ or NRR′;

each R is independently selected from H, alkyl or haloalkyl;R′ is H, alkyl, NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂;CR₂C(═O)NR₂; C(═NR)R; C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; orC(═O)CR₂NRC(═NR)NR₂; andZ is H, alkyl, halo, CN, OR, SR′ or NRR′.

In one embodiment, R¹ is H. In one embodiment, R¹ is alkyl, for exampleCH₃. In one embodiment, R² is H. In one embodiment, R² is alkyl, forexample CH₃. In one embodiment, both R¹ and R² are alkyl, for exampleCH₃.

In one embodiment, P is H. In one embodiment, P is OH. In oneembodiment, P is halogen. In one embodiment, P is hydroxyl protected bya hydroxyl protecting group. In a particular embodiment, P is OH orhydroxyl protected by a hydroxyl protecting group.

In one embodiment, X is —(CR₂)_(m)—. In one subembodiment, m is 0. Inanother subembodiment, m is 1. In another subembodiment, m is 2. In oneembodiment, X is —C(═O)—.

In one embodiment, at least one R is H. In one embodiment, at least twoRs are H. In one embodiment, at least one R is alkyl, for example CH₃,CH₂CH₃ or CH₂CH₂CH₃. In one embodiment, at least one R is haloalkyl, forexample CF₃.

In one embodiment, Y is CN. In another embodiment, Y is OR. In aparticular embodiment, Y is OH. In one embodiment, Y is SR′, for exampleSH or S(alkyl). In one embodiment, Y is SR′ and R′ is C(═NR)NR₂, forexample C(═NH)NH₂, C(═NCH₃)NH₂, C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃)₂,C(═NR)NH(CH₃), or C(═NCF₃)NH₂. In one embodiment, Y is NRR′. In asubembodiment, Y is NHR′. In another subembodiment, Y is N(alkyl)R′, forexample N(CH₃)R′.

In one embodiment, Z is H. In one embodiment, Z is halogen. In anotherembodiment, Z is alkyl. In one embodiment, Z is CN. In anotherembodiment, Z is halo. In certain embodiments, Z is OR, for example OH.In one embodiment, Z is SR′, for example SH or S(alkyl). In oneembodiment, Z is halogen, for example mono- or multi-F or Cl. In oneembodiment, Z is NRR′. In a subembodiment, Z is NHR′. In anothersubembodiment, Z is N(alkyl)R′, for example N(CH₃)R′.

In one embodiment, R′ is H. In one embodiment, R′ is alkyl, for exampleCH₃. In one embodiment, R′ is NR₂, for example NH₂, NHR, NHCH₃, orN(CH₃)₂. In another embodiment, R′ is C(═O)R, for example C(═O)CH₃ orC(═O)CF₃. In another embodiment, R′ is SO₂R, for example SO₂CH₃. Inanother embodiment, R′ is SO₂NR₂, for example SO₂NH₂. In anotherembodiment, R′ is C(═NR)NR₂, for example C(═NH)NH₂, C(═NCH₃)NH₂,C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃)₂, C(═NR)NH(CH₃), or C(═NCF₃)NH₂. Inanother embodiment, R′ is C(═O)NR₂, for example C(═O)NH₂, C(═O)NHR,C(═O)NHCH₃ or C(═O)N(CH₃)₂. In another embodiment, R′ is C(═NR)R, forexample C(═NH)H, C(═NH)R or C(═NH)CH₃. In another embodiment, R′ isC(═NR)NRSO₂R, for example C(═NH)NHSO₂H, C(═NH)NHSO₂R, or C(═NH)NHSO₂CH₃.In another embodiment, R′ is C(═NR)NRC(═O)R, for example C(═NH)NHC(═O)Ror C(═NH)NHC(═O)CH₃. In another embodiment, R′ is C(═O)CR₂NRSO₂NR₂, forexample C(═O)CH₂NHSO₂NR₂, C(═O)CH₂NHSO₂NH₂, or C(═O)CH₂NHSO₂N(CH₃)₂. Inanother embodiment, R′ is C(═O)CR₂NRC(═NR)NR₂, for exampleC(═O)CH₂NHC(═NH)NR₂, C(═O)CH₂NHC(═NH)NH₂ or C(═O)CH₂NHC(═NH)N(CH₃)₂.

In one embodiment, the compound of Formula III is selected from thegroup consisting of:

In another particular embodiment, the compound is a compound of FormulaIV,

or a pharmaceutically acceptable salt, ester or prodrug thereof,whereinR¹, R² and R³ are each independently selected from H or alkyl;P is H, OH, halogen, or hydroxyl protected by a hydroxyl protectinggroup;n is 0, 1, or 2X is —(CR₂)_(m)— or —C(═O)—;m is 0, 1 or 2;

Y is CN, OR, SR′ or NRR′;

each R is independently selected from H, alkyl or haloalkyl;R′ is H, alkyl, NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂;CR₂C(═O)NR₂; C(═NR)R; C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; orC(═O)CR₂NRC(═NR)NR₂; andZ is H, alkyl, halo, CN, OR, SR′ or NRR′.

In one embodiment, R¹ is H. In one embodiment, R¹ is alkyl, for exampleCH₃. In one embodiment, R² is H. In one embodiment, R² is alkyl, forexample CH₃. In one embodiment, both R¹ and R² are alkyl, for exampleCH₃.

In one embodiment, P is H. In one embodiment, P is OH. In oneembodiment, P is halogen. In one embodiment, P is hydroxyl protected bya hydroxyl protecting group. In a particular embodiment, P is OH orhydroxyl protected by a hydroxyl protecting group.

In one embodiment, n is 0. In one embodiment, n is 1. In anotherembodiment, n is 2. In one embodiment, n is 1 or 2. In one embodiment, nis 0.

In one embodiment, X is —(CR₂)_(m)—. In one subembodiment, m is 0. Inanother subembodiment, m is 1. In another subembodiment, m is 2. In oneembodiment, X is —C(═O)—.

In one embodiment, at least one R is H. In one embodiment, at least twoRs are H. In one embodiment, at least one R is alkyl, for example CH₃,CH₂CH₃ or CH₂CH₂CH₃. In one embodiment, at least one R is haloalkyl, forexample CF₃.

In one embodiment, Y is CN. In another embodiment, Y is OR. In aparticular embodiment, Y is OH. In one embodiment, Y is SR′, for exampleSH or S(alkyl). In one embodiment, Y is SR′ and R′ is C(═NR)NR₂, forexample C(═NH)NH₂, C(═NCH₃)NH₂, C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃)₂,C(═NR)NH(CH₃), or C(═NCF₃)NH₂. In one embodiment, Y is NRR′. In asubembodiment, Y is NHR′. In another subembodiment, Y is N(alkyl)R′, forexample N(CH₃)R′.

In one embodiment, R′ is H. In one embodiment, R′ is alkyl, for exampleCH₃. In one embodiment, R′ is NR₂, for example NH₂, NHR, NHCH₃, orN(CH₃)₂. In another embodiment, R′ is C(═O)R, for example C(═O)CH₃ orC(═O)CF₃. In another embodiment, R′ is SO₂R, for example SO₂CH₃. Inanother embodiment, R′ is SO₂NR₂, for example SO₂NH₂. In anotherembodiment, R′ is C(═NR)NR₂, for example C(═NH)NH₂, C(═NCH₃)NH₂,C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃)₂, C(═NR)NH(CH₃), or C(═NCF₃)NH₂. Inanother embodiment, R′ is C(═O)NR₂, for example C(═O)NH₂, C(═O)NHR,C(═O)NHCH₃ or C(═O)N(CH₃)₂. In another embodiment, R′ is C(═NR)R, forexample C(═NH)H, C(═NH)R or C(═NH)CH₃. In another embodiment, R′ isC(═NR)NRSO₂R, for example C(═NH)NHSO₂H, C(═NH)NHSO₂R, or C(═NH)NHSO₂CH₃.In another embodiment, R′ is C(═NR)NRC(═O)R, for example C(═NH)NHC(═O)Ror C(═NH)NHC(═O)CH₃. In another embodiment, R′ is C(═O)CR₂NRSO₂NR₂, forexample C(═O)CH₂NHSO₂NR₂, C(═O)CH₂NHSO₂NH₂, or C(═O)CH₂NHSO₂N(CH₃)₂. Inanother embodiment, R′ is C(═O)CR₂NRC(═NR)NR₂, for exampleC(═O)CH₂NHC(═NH)NR₂, C(═O)CH₂NHC(═NH)NH₂ or C(═O)CH₂NHC(═NH)N(CH₃)₂.

In one embodiment, Y is CN, NR₂, SC(═NR)NR₂, C(═O)NR₂, C(═-O)NRSO₂R;C(═O)NRSO₂NR₂; NRC(═NR)NR₂; NRSO₂NR₂; NRC(O)NR₂; NRCR₂C(O)NR₂;NRCR(═NR); CR₂NRC(═NR)NR₂; NRC(═NR)NRSO₂R; NRC(═NR)NRC(O)R;C(O)NRCR₂C(O)NR₂; C(O)NRC(═NR)NR₂; OR; NRC(O)CR₂NRSO₂NR₂; orNRC(O)CR₂NRC(═NR)NR₂.

In one embodiment, Z is H. In one embodiment, Z is halogen. In anotherembodiment, Z is alkyl. In one embodiment, Z is CN. In anotherembodiment, Z is halo. In certain embodiments, Z is OR, for example OH.In one embodiment, Z is SR′, for example SH or S(alkyl). In oneembodiment, Z is halogen, for example mono- or multi-F or Cl. In oneembodiment, Z is NRR′. In a subembodiment, Z is NHR′. In anothersubembodiment, Z is N(alkyl)R′, for example N(CH₃)R′.

In one embodiment, R¹ is alkyl; R² is alkyl; P is hydroxyl or hydroxylprotected by a hydroxyl protecting group; n is 0, 1 or 2; m is 0; and Yis —CN. In another embodiment, R¹ is alkyl; R² is alkyl; P is hydroxylor hydroxyl protected by a hydroxyl protecting group; n is 0, 1 or 2; mis 0; and Y is OR. In another embodiment, R¹ is alkyl; R² is alkyl; P ishydroxyl or hydroxyl protected by a hydroxyl protecting group; n is 0, 1or 2; m is 0 or 1; and Y is NRR′.

In one embodiment, when Y is CN, X is not —C(═O)—. In anotherembodiment, R¹ is alkyl; R² is alkyl; P is hydroxyl or hydroxylprotected by a hydroxyl protecting group; n is 0, 1 or 2; X is —C(═O)—;and Y is OR. In another embodiment, R¹ is alkyl; R² is alkyl; P ishydroxyl or hydroxyl protected by a hydroxyl protecting group; n is 0, 1or 2; X is —C(═O)—; and Y is NRR′.

In one embodiment, R¹ is alkyl; R² is alkyl; P is hydroxyl or hydroxylprotected by a hydroxyl protecting group; n is 1 or 2; m is 0; and Y is—CN. In another embodiment, R¹ is alkyl; R² is alkyl; P is hydroxyl orhydroxyl protected by a hydroxyl protecting group; n is 1 or 2; m is 0;and Y is OR. In another embodiment, R¹ is alkyl; R² is alkyl; P ishydroxyl or hydroxyl protected by a hydroxyl protecting group; n is 1 or2; m is 0 or 1; and Y is NRR′. In one embodiment, R¹ is alkyl; R² isalkyl; P is hydroxyl or hydroxyl protected by a hydroxyl protectinggroup; n is 1 or 2; X is —C(═O)—; and Y is OR. In another embodiment, R¹is alkyl; R² is alkyl; P is hydroxyl or hydroxyl protected by a hydroxylprotecting group; n is 1 or 2; X is —C(═O)—; and Y is NRR′.

In another particular embodiment, the compound is a compound of FormulaV,

or a pharmaceutically acceptable salt, ester or prodrug thereof,whereinR¹, R² and R³ are each independently selected from H or alkyl;P is H, OH, halogen, or hydroxyl protected by a hydroxyl protectinggroup;X is —(CR₂)_(m)— or —C(═O)—;m is 0, 1 or 2;

Y is CN, OR, SR′ or NRR′;

each R is independently selected from H, alkyl or haloalkyl;R′ is H, alkyl, NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂;CR₂C(═O)NR₂; C(═NR)R; C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; orC(═O)CR₂NRC(═NR)NR₂; andZ is H, alkyl, halo, CN, OR, SR′ or NRR′.

In one embodiment, R¹ is H. In one embodiment, R¹ is alkyl, for exampleCH₃. In one embodiment, R² is H. In one embodiment, R² is alkyl, forexample CH₃. In one embodiment, both R¹ and R² are alkyl, for exampleCH₃.

In one embodiment, P is H. In one embodiment, P is OH. In oneembodiment, P is halogen. In one embodiment, P is hydroxyl protected bya hydroxyl protecting group. In a particular embodiment, P is OH orhydroxyl protected by a hydroxyl protecting group.

In one embodiment, X is —(CR₂)_(m)—. In one subembodiment, m is 0. Inanother subembodiment, m is 1. In another subembodiment, m is 2. In oneembodiment, X is —C(═O)—.

In one embodiment, at least one R is H. In one embodiment, at least twoRs are H. In one embodiment, at least one R is alkyl, for example CH₃,CH₂CH₃ or CH₂CH₂CH₃. In one embodiment, at least one R is haloalkyl, forexample CF₃.

In one embodiment, Z is H. In one embodiment, Z is halogen. In anotherembodiment, Z is alkyl. In one embodiment, Z is CN. In anotherembodiment, Z is halo. In certain embodiments, Z is OR, for example OH.In one embodiment, Z is SR′, for example SH or S(alkyl). In oneembodiment, Z is halogen, for example mono- or multi-F or Cl. In oneembodiment, Z is NRR′. In a subembodiment, Z is NHR′. In anothersubembodiment, Z is N(alkyl)R′, for example N(CH₃)R′.

In one embodiment, Y is CN. In another embodiment, Y is OR. In aparticular embodiment, Y is OH. In one embodiment, Y is SR′, for exampleSH or S(alkyl). In one embodiment, Y is SR′ and R′ is C(═NR)NR₂, forexample C(═NH)NH₂, C(═NCH₃)NH₂, C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃)₂,C(═NR)NH(CH₃), or C(═NCF₃)NH₂. In one embodiment, Y is NRR′. In asubembodiment, Y is NHR′. In another subembodiment, Y is N(alkyl)R′, forexample N(CH₃)R′.

In one embodiment, R′ is H. In one embodiment, R′ is alkyl, for exampleCH₃. In one embodiment, R′ is NR₂, for example NH₂, NHR, NHCH₃, orN(CH₃)₂. In another embodiment, R′ is C(═O)R, for example C(═O)CH₃ orC(═O)CF₃. In another embodiment, R′ is SO₂R, for example SO₂CH₃. Inanother embodiment, R′ is SO₂NR₂, for example SO₂NH₂. In anotherembodiment, R′ is C(═NR)NR₂, for example C(═NH)NH₂, C(═NCH₃)NH₂,C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃) 2, C(═NR)NH(CH₃), or C(═NCF₃)NH₂. Inanother embodiment, R′ is C(═O)NR₂, for example C(═O)NH₂, C(═O)NHR,C(═O)NHCH₃ or C(═O)N(CH₃)₂. In another embodiment, R′ is C(═NR)R′, forexample C(═NH)H, C(═NH)R or C(═NH)CH₃. In another embodiment, R′ isC(═NR)NRSO₂R, for example C(═NH)NHSO₂H, C(═NH)NHSO₂R, or C(═NH)NHSO₂CH₃.In another embodiment, R′ is C(═NR)NRC(═O)R, for example C(═NH)NHC(═O)Ror C(═NH)NHC(═O)CH₃. In another embodiment, R′ is C(═O)CR₂NRSO₂NR₂, forexample C(═O)CH₂NHSO₂NR₂, C(═O)CH₂NHSO₂NH₂, or C(═O)CH₂NHSO₂N(CH₃)₂. Inanother embodiment, R′ is C(═O)CR₂NRC(═NR)NR₂, for exampleC(═O)CH₂NHC(═NH)NR₂, C(═O)CH₂NHC(═NH)NH₂ or C(═O)CH₂NHC(═NH)N(CH₃)₂.

In one embodiment, the compound of Formula IV is the compound:

In another particular embodiment, the compound is a compound of FormulaVI,

or a pharmaceutically acceptable salt, ester or prodrug thereof,whereinR¹ and R² are each independently selected from H or alkyl;P is H, OH, halogen, or hydroxyl protected by a hydroxyl protectinggroup;X is —(CR₂)_(m)— or —C(═O)—;m is 0, 1 or 2;

Y is CN, OR, SR′ or NRR′;

each R is independently selected from H, alkyl or haloalkyl;R′ is H, alkyl, NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂;CR₂C(═O)NR₂; C(═NR)R; C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; orC(═O)CR₂NRC(═NR)NR₂; andZ is H, alkyl, halo, CN, OR, SR′ or NRR′.

In one embodiment, R¹ is H. In one embodiment, R¹ is alkyl, for exampleCH₃. In one embodiment, R² is H. In one embodiment, R² is alkyl, forexample CH₃. In one embodiment, both R¹ and R² are alkyl, for exampleCH₃.

In one embodiment, P is H. In one embodiment, P is OH. In oneembodiment, P is halogen. In one embodiment, P is hydroxyl protected bya hydroxyl protecting group. In a particular embodiment, P is OH orhydroxyl protected by a hydroxyl protecting group.

In one embodiment, X is —(CR₂)_(m)—. In one subembodiment, m is 0. Inanother subembodiment, m is 1. In another subembodiment, m is 2. In oneembodiment, X is —C(═O)—.

In one embodiment, at least one R is H. In one embodiment, at least twoRs are H. In one embodiment, at least one R is alkyl, for example CH₃,CH₂CH₃ or CH₂CH₂CH₃. In one embodiment, at least one R is haloalkyl, forexample CF₃.

In one embodiment, Y is CN. In another embodiment, Y is OR. In aparticular embodiment, Y is OH. In one embodiment, Y is SR′, for exampleSH or S(alkyl). In one embodiment, Y is SR′ and R′ is C(═NR)NR₂, forexample C(═NH)NH₂, C(═NCH₃)NH₂, C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃)₂,C(═NR)NH(CH₃), or C(═NCF₃)NH₂. In one embodiment, Y is NRR′. In asubembodiment, Y is NHR′. In another subembodiment, Y is N(alkyl)R′, forexample N(CH₃)R′.

In one embodiment, Z is H. In one embodiment, Z is halogen. In anotherembodiment, Z is alkyl. In one embodiment, Z is CN. In anotherembodiment, Z is halo. In certain embodiments, Z is OR, for example OH.In one embodiment, Z is SR′, for example SH or S(alkyl). In oneembodiment, Z is halogen, for example mono- or multi-F or Cl. In oneembodiment, Z is NRR′. In a subembodiment, Z is NHR′. In anothersubembodiment, Z is N(alkyl)R′, for example N(CH₃)R′.

In one embodiment, the Z substituent and the X—Y substituent are in acis-configuration with respect to each other. In another embodiment, theZ substituent and the X—Y substituent are in a trans-configuration withrespect to each other. In a preferred embodiment, Z is hydroxyl. Inanother preferred embodiment, m is 0 and Y is NRR′, where R is H and R′is C(═NR)NR₂. In yet another preferred embodiment, Z is hydroxyl, m is 0and Y is NRR′, where R is H and R′ is C(═NR)NR₂.

In one embodiment, R′ is H. In one embodiment, R′ is alkyl, for exampleCH₃. In one embodiment, R′ is NR₂, for example NH₂, NHR, NHCH₃, orN(CH₃)₂. In another embodiment, R′ is C(═O)R, for example C(═O)CH₃ orC(═O)CF₃. In another embodiment, R′ is SO₂R, for example SO₂CH₃. Inanother embodiment, R′ is SO₂NR₂, for example SO₂NH₂. In anotherembodiment, R′ is C(═NR)NR₂, for example C(═NH)NH₂, C(═NCH₃)NH₂,C(═NCH₃)NHCH₃, C(═NCH₃)N(CH₃) 2, C(═NR)NH(CH₃), or C(═NCF₃)NH₂. Inanother embodiment, R′ is C(═O)NR₂, for example C(═O)NH₂, C(═O)NHR,C(═O)NHCH₃ or C(═O)N(CH₃)₂. In another embodiment, R′ is C(═NR)R, forexample C(═NH)H, C(═NH)R or C(═NH)CH₃. In another embodiment, R′ isC(═NR)NRSO₂R, for example C(═NH)NHSO₂H, C(═NH)NHSO₂R, or C(═NH)NHSO₂CH₃.In another embodiment, R′ is C(═NR)NRC(═O)R, for example C(═NH)NHC(═O)Ror C(═NH)NHC(═O)CH₃. In another embodiment, R′ is C(═O)CR₂NRSO₂NR₂, forexample C(═O)CH₂NHSO₂NR₂, C(═O)CH₂NHSO₂NH₂, or C(═O)CH₂NHSO₂N(CH₃)₂. Inanother embodiment, R′ is C(═O)CR₂NRC(═NR)NR₂, for exampleC(═O)CH₂NHC(═NH)NR₂, C(═O)CH₂NHC(═NH)NH₂ or C(═O)CH₂NHC(═NH)N(CH₃)₂.

In one embodiment, the compound of Formula VI is selected from the groupconsisting of:

In certain embodiments, the compound is selected from the groupconsisting of:

Method of Treatment

The present invention also provides a method of preventing or treating abacterial infection, in a host, for example an animal, and typically ahuman, including administering a therapeutic amount of a compound of thepresent invention, or a pharmaceutically acceptable salt and/or prodrugtherein, optionally in a pharmaceutically acceptable carrier or diluentwhere the bacterial infection is due to gram-negative bacteria. In oneembodiment, the bacterial infection is a drug resistant and/ormultiple-drug resistant bacterial infection.

The invention also provides a compound of the present invention for usein medical therapy.

The present invention also provides a use of a therapeutic amount of acompound of the present invention, or a pharmaceutically acceptable saltand/or prodrug therein, optionally in a pharmaceutically acceptablecarrier or diluent, for preventing or treating a gram-negative bacterialinfection, in a host, such as an animal, and typically a human.

The distinctive feature of gram-negative bacteria is the presence of adouble membrane surrounding each bacterial cell. Although all bacteriahave an inner cell membrane, gram-negative bacteria have a unique outermembrane. This outer membrane excludes certain drugs and antibioticsfrom penetrating the cell, partially accounting for why gram-negativebacteria are generally more resistant to antibiotics than aregram-positive bacteria. The pathogenic capability of gram-negativebacteria is usually associated with certain components of their cellwalls, particularly the lipopolysaccharide (endotoxin) layer.

The outer membrane of gram-negative bacteria is rich inlipopolysaccharide. If gram-negative bacteria enter the bloodstream,lipopolysaccharide can trigger a cascade of events, including high feverand a drop in blood pressure. Unlike Gram-positive bacteria, whichassume a violet color in Gram staining, Gram-negative bacteriaincorporate the counterstain rather than the primary stain. Because thecell wall of Gram (−) bacteria is high in lipid content and low inpeptidoglycan content, the primary crystal-violet escapes from the cellwhen the decolorizer is added. Most enteric (bowel related) illnessescan also be attributed to this group of bacteria.

Examples of gram-negative bacteria include Aeromonas sp., Acinetobactersp. such as Acinetobacter baumannii(or A. calcoaceticus), Actinobacillusactinomycetemcomitans, Bacteroides sp. such as Bacteroides fragilis,Bartonella, Bdellovibrio spp., Bordetella pertussis, Brucella sp.,Burkholderia cepacia, Burkholderia, pseudomallei, Campylobacter sp.,Capnocytophaga sp., Cardiobacterium hominis, Chlamydia trachomatis,Citrobacter sp., Eikenella corrodens, Enterobacter sp., Escherichiacoli, Francisella tularensis, Flavobacterium sp., Fusobacterium sp.,Helicobacter pylori, Haemophilus influenzae, Haemophilus ducreyi,Klebsiella spp. such as Klebsiella pneumoniae, Kingella kingae,Legionella spp. such as Legionella pneumophila, Moraxella catarrhalis,Morganella, Neisseria gonorrhoeae, Neisseria meningitidis, Pasteurellapestis, Pasteurella multocida, Plesiomonas shigelloides, Prevotella sp.,Proteus spp., Providencia, Pseudomonas spp. such as Pseudomonasaeruginosa, Salmonella spp. such as Salmonella enteriditis andSalmonella typhi, Serratia marcescens, Shigella spp., Vibrio cholerae,Vibrio parahaemolyticus, Vibrio vulnificus, Veillonella sp., Xanthomonasmaltophilia or Stenotrophomonas maltophila, Yersinia pestis, Yersiniaenterocolitica. Additionally, some organisms simply tend not to be welldifferentiated by gram staining, despite any known phylogeneticaffiliation with the gram-negatives or gram-positives. Rickettsiaprowazekii, Rickettsia rickettsii and Treponema pallidum. Chlamydias aresmall, gram-negative, peptidoglycan-less cocci that are obligateintracellular parasites of animals. Spirochetes are chemoheterotrophicbacteria whose cells are tightly coiled or resemble a stretched springwith gram-negative-like cell envelopes. Spirochetes include Spirillumminus, Borrelia burgdorferi (Lyme disease), Leptospira spp.(leptospirosis) and Treponema pallidum (syphilis). Rickettsias andactinomycetes are also gram-negative pleomorphic bacilli andcoccobacilli that are obligate intracellular parasites of eucaryotestransmitted generally by insects and ticks.

The present invention also provides a use of a therapeutic amount of acompound of the present invention, or a pharmaceutically acceptable saltand/or prodrug therein, optionally in a pharmaceutically acceptablecarrier or diluent, in the manufacture of a medicament for preventing ortreating a gram-negative bacterial infection, in a host, such as ananimal, and typically a human.

The invention also includes methods of inhibiting bacterial infection ina host. Inhibition of bacterial replication or treatment of an infectionin a cell can be measured by showing a reduction in bacterialreplication in a cell to a level lower than the level in an otherwiseidentical cell, which was not administered the compound of theinvention. The reduction can be by about 80%, 85%, 90%, 95%, about 99.9%or more. The level of bacterial replication in a cell can be assessed byany known methods. For example, the level of bacterial replication in acell can be assessed by evaluating the number of bacterial particles oramount of a bacterial component, such as a bacterial protein, abacterial enzyme, or bacterial nucleic acid, in the cell or in fluid ordebris associated with the cell. The number of infectious bacteria in acell can be evaluated, for example, in a plaque assay. The level of abacterial component such as a bacterial protein or enzyme in a cell canbe evaluated using standard analytical techniques of proteinbiochemistry, such as, for example, using an activity assay for abacterial enzyme, or using Western blotting or quantitative gelelectrophoresis for a bacterial protein. Bacterial nucleic acid levelsin a cell can be evaluated using standard analytical techniques such asNorthern blotting and Southern Blotting or quantitation by polymerasechain reaction (PCR).

As used herein, to inhibit bacterial replication in a host means toreduce the bacterial load in a host to a level, which is lower than thelevel of the bacterial load in an otherwise identical host, which wasnot administered the compound. Bacterial load in a mammal can be reducedby about 1 to 12 log₁₀ or more relative to an otherwise identicalmammal, which was not administered the compound. Bacterial load in amammal can be assessed by a number of methods known in the art such as,for example, obtaining a tissue or fluid sample from the mammal andassessing the amount of bacterial components in the mammal containedtherein using technology which is either immunological, biochemical ormolecular biological in nature and which is well known to the skilledartisan and which are described elsewhere herein. Inhibition ofbacterial replication in a cell is assessed using similar or identicalassays as those used to assess bacterial load in a mammal.

Combination and Alternation Therapies

In one embodiment of the invention, one or more therapeutic agents,including particularly antimicrobial agents such as antibiotic agentsthat are effective against gram negative bacteria, can be used incombination and/or alternation with the compound/composition of thepresent invention to achieve a additive and/or synergistic therapeuticeffect.

The active compounds can be administered in combination, alternation orsequential steps with another anti-bacterial agent. In combinationtherapy, effective dosages of two or more agents are administeredtogether, whereas in alternation or sequential-step therapy, aneffective dosage of each agent is administered serially or sequentially.The dosages given will depend on absorption, inactivation and excretionrates of the drug as well as other factors known to those of skill inthe art. It is to be noted that dosage values will also vary with theseverity of the condition to be alleviated. It is to be furtherunderstood that for any particular subject, specific dosage regimens andschedules should be adjusted over time according to the individual needand the professional judgment of the person administering or supervisingthe administration of the compositions. In some embodiments, ananti-bacterial agent exhibits an EC₅₀ of 10-15 μM or less, or typicallyless than 1-5 μM.

In one particular embodiment, the combination includes a β-lactamaseinhibitor, such as clavulanic acid, which has been used as in thedelivery of prophylactic amounts of antibiotics in patients. Althoughclavulanic acid does have some degree of bacterial activity, itsprincipal role is as a beta-lactamase inhibitor. Clavulanic acid has asimilar structure to the beta-lactam antibiotics but binds irreversiblyto the beta-lactamase enzymes. Used in combination with the beta-lactamantibiotics, it has become one of the most prescribed antibiotics in thewestern world prolonging the effective life of antibiotics such asAmpicillin (as in GSK's Augmentin®).

It is possible that drug-resistant variants of bacteria can emerge afterprolonged treatment with an anti-bacterial agent. The efficacy of a drugagainst the bacterial infection can be prolonged, augmented, or restoredby administering the compound in combination or alternation with asecond, and perhaps third, anti-bacterial agent, for example with adifferent site of activity than the principle drug. Alternatively, thepharmacokinetics, biodistribution or other parameter of the drug can bealtered by such combination or alternation therapy.

Suitable antibiotic agents are disclosed, e.g. in Physician's Desk 30Reference (PDR), Medical Economics Company (Montvale, N.J.), (53rd Ed.),1999; Mayo Medical Center Formulary, Unabridged Version, Mayo Clinic(Rochester, Minn.), January 1998; Merck Index An Encyclopedia ofChemicals, Drugs and Biologicals, (11th Ed.), Merck & Co., Inc. (Rahway,N.J.), 1989; University of Wisconsin Antimicrobial Use Guide,http://www.medsch.wisc.edu/clinsci/5amcg/amcg.html; Introduction on theUse of the Antibiotics Guideline, of Specific Antibiotic Classes, ThomasJefferson University,http://jeffiine.tju.edu/CWIS/OAC/antibiotics_guide/intro.html; andreferences cited therein.

Nonlimiting examples of agents that can be used in combination oralternation with the compounds of the invention include:aminoglycosides, β-lactam antibiotics, cephalosporius, macrolides,miscellaneous antibiotics, penicillins, tetracyclines, antifungals,antimalarial agents, antituberculosis agents, antibacterials,leprostatics, miscellaneous anti-infectives, quinolones, sulfonamides,urinary anti-infectives, nasal antibiotics, opthalmic antibiotics,opthalmic antibacterials, opthalmicquinalones, opthalmic sulfonamides,skin and mucous membrane antibiotics, skin and mucous membraneantifungals, skin and mucous membrane antibacterials, skin and mucousmembrane miscellaneous anti-infectives, skin and mucousmembranescabicides and pedulicides, skin and mucous membraneantincoplasts, nitrofurans and oxazolidinones.

Specific compounds include, for example, Amikacin (amikacin sulfate);Craramyein (gentamicin sulfate); Nebcin (tobramycin sulfate); Netromycin(netilmicin sulfate); Streptomycin Sulfate; and TOBI (tobramycin),Azactam (aztreonam); Cefotan (cefotetan); Lorabid (loracarbef); Mefoxin(cefoxitin); Merrem (meropenem); and Primaxin (imipenem and cilastatinfor injectable suspension); Ancef (cefazolin); Ceclor (cefaclor); Cedax(ceffibuten); Cefizox (ceffizoxime sodium); Cefobid (cefoperazonesodium); Ceftin (cefuroxime axetil); Cefzil (cefprozil); Ceptaz(ceftazidime); Claforan (cefotaxime); Duricef (cefadroxil monohydrate);Fortaz (ceftazidime); Keflex (cephalexin); Keftab (cephalexin HCl);Kefurox (cefuroxime); Kcfzol (ccfazolin); Mandol (cefamandole nafatc);Maxipimc (cefepime HCl); Monocid (cefonicidsodium); Omnicef (cefdinir);Rocephin (ceftriaxone); Suprax (cefixime); Tazicef (ceftazidime);Tazidime (ceftazidime); Vantin (cefpodoxime proxetil); andZinacef5(cefuroxime); Biaxin (clarithromycin); Dynabac (dirithromycin);E.E.S. 200 (Erythromycin Ethylsuccinate); E.E.S. 400 (ErythromycinEthylsuccinate); Ery-Ped 200 (Erythromycin Ethylsuccinate); EryPed 400(Erythromycin Ethylsuccinate); Ery-Tab (Erythromycin delayed-releasetablets); Erythrocin Stearate (Erythromycin stearate); Ilosone(erythromycinestolate); PCE Dispertab (erythromycin particles intablets); Pediazole (erythromycin ethylsuccinate and sulfisoxazoleacetyl for oral suspension); Tao (troleandomycin); Zithromax(azithromycin); and Erythromycin; Cleocin HCl (clindamycinhydrochloride); Cleotin Phosphate (elindamycin phosphate); Coly-Mycin M(colistimethate sodium); and Vancocin HCl (vancomycin hydrochloride);Amoxil (amoxicillin); Augmentin (amoxicillin/clavulanate potassium);Bicillin C-R 900/300 (Penicillin G benzathine and Penicillin G procainesuspension); Bicillin C-R (Penicillin G benzathine and Penicillin Gprocaine suspension); Bicillin L-A (Penicillin G benzathine suspension);Geoeillin (carbencillin indanyl sodium); Mezlin (sterilemezlocillinsodium); Omnipen (ampicillin); Pen-Vee K (penicillin Vpotassium); Pfizerpen (penicillin G potassium); Pipracil (piperacillinsodium); Speetrobid (bacampicillin-HCl); Ticar (tiearcillin disodium);Timentin (ticarcillin disodium and clavulanate potassium); Unasyn(ampicillin sodium/sulbactam sodium); Zosyn (piperacillin sodium andtazobactam sodium); and Dicloxacillin Sodium; Achromycin V (tetracyclineHCl); Declomycin (derneclo-cycline HCl); Dynacin (minocylcine HCl);Minocin (minocyclinc hydrochloride); Monodox (Doxycycline monohydratecapsules); Terramycin (oxytetracyline); Vectrin (minocyclinehydrochloride); Vibramycin Calcium (doxycycline sodium); VibramycinHyclate (doxycycline hyclate); Vibramycin Monohydrate (doxycyclinemonohydrate); Vibra-Tabs (doxycycline-hydrate); Declomycin(demeclocycline HCl); Vibramycin (doxycycline); Dynacin (MinocylineHCl); Terramycin (oxytetracycline HCl); Achromycin V capsules5(tetracycline HCl); Linco-mycins; and Cleotin HCl (clindamycin HCl);Abelcet (amphotericin B lipid complex); AmBisome (amphotericin B);Amphotec (amphotericin B cholesterol sulfatecomplex); Ancobon(flucytosine); Diflucan (fluconazole); Fulvicin P/Gamma (ultramicrosizegriseofulvin); Fulvicin P/G 165 and 330 (ultramicrosize griseofulvin);Grifulvin V (griseofulvin); Gals-PEG (gxiseofulvin ultramicrosize);Lamisil (terbinafine hydrochloride); Nizoral (ketoconazole);Amphotericin B; Lotrimin (clotrimazole); Dapsone tablets (dapsone);Diflucan (fluconazole); Monistat-Derm cream (miconazole); Mycostalin Crc.am (nystatin); and Sporanox (itraconazole); Aralen hydrochloride(chloroquine HCl); Aralen phosphate (chloroquine phosphatc); Dataprim(pyrimethamine); Ladam (mefloquine HCl); and Plaquenil(hydroxychloroqnine sulfate); Capastat sulfate (capreomycinsulfate);Myambutol (ethambutol hydrochloride); Mycobutin (rifabutin capsules);Nydrazid (isoniazid injection); Paser (aminosalicylic acid); Prifiin(rifapentine); Pyrazinamide tablets (pyrazinamide); Rifadin (rifampincapsules); Rifadin IV (rifampin for injection); Rifamate (rifampin andisoniazid); Rifater (rifampin, isoniazid and pyrazinamide); Seromycin(cycloserine capsules); Streptomycin-Sulfate; Tice BCG (BCG vaccine);Cycloserine (seromycin capsules); Urised (Methenamine); and Trecator-SC(ethionamide tablets); Alferon N (interferon alfa-n3); Crixivan(indinavir sulfate); Cytovene (ganciclovir); Cytovene-IV (ganciclovirsodium); Epivir (lamivudine); Famvir (famciclovir); Flumadine(rimantadine HCl); Foscavir (foscamet sodium); Hivid (zalcitabine);Intron A (interferon alfa-2b); Invirase (saquinavir mesylate); Norvir(ritonavir); Rebetron combination therapy, which contains Rebetrol(ribavirin) and Intron A (inteferon alfa-2b); Rescriptor (delavirdinemesylate); Retrovir (ziduvudine); Retrovir IV (ziduvudine); Symmetrel(amantadine HCl); Synagis (palivizumab); Valtrex (valacyclovir HCl);Videx (didanosine); Viracept (nelfinavir mesylate); Viramune(nevirapine); Virazole (ribavirin); Vistide (cidofovir); Zerit(stavudine (d4T)); Symmetrel Syrup (amantadine HCl); Combivir Tablets(lamiduvine); and Zovirax (acyclovir); Dapsone Tablets (dapsone);Daraprim (pyrimethamine); Flagyl 375 (metronidazole); Flagyl ER Tablets(metronidazole); Flagyl I.V. (metronidazole); Furoxone (furazolidone);Mepron (atovaquone); and Neutrexin (tfirnetrexate glucuronate); Cipro(ciprofloxacin HCl); Floxin (ofloxacin); Levaquin (levofloxacin);Mazaquin (lomcfioxacin HCl); Noroxin (norfloxacin); Penetrex (enoxacin);Raxar (grepafloxacin HCl); Trovan (trovafioxacin mesylate); and Zagam(sparfloxacin); Bactrim. (trimethoprim and sulfamethoxazole); Bactrim DS(Irimethoprim and sulfamethoxazole double strength); Pediazole(erythromycin ethylsuccinate and sulfisoxazole acetyl); Septra(trimethoprim and sulfamethoxazole); Septra DS (trimethoprim andsulfamethoxazole); Co-Trimoxazole, Sulfadiazine, Battrim I.V. Infusion(sulfamethoxazole); Sulfapyridine and Pediazole (erythromycinethylsuccinate and sulfisoxazole acetyl); Furadantin (nitrofurantoin);Macrobid (nitrofurantoin monohydrate macrocrystals); Macrodantin(nitrofurantoin macrocrystals); Monurol Sachet (fosfomycintromethamine); NegGram Caplets (nalidixic acid); Septra (trimethoprimand sulfamethoxazole); Septra DS(trimethoprim and sulfamethoxazole);Urised (a combination of the antisepticsmethenamine, methylene blue,phenyl salicylate, benzoic acid and parasympatholytics (atropinesulfate) hyoscyamine); (oxytetracyclinc HCl, sulfamethizole andphenazopyridine HCl); (methenaminc mandelatc); Bactroban (mupirocin);Chloromycetin opthalmic (chloramphenical); Cortisporin (neomycin andpolymyxin [3 sulfates and hydrocortisone acetate cream); Ilotycin(erythromycin opthalmic ointment); NeoDecadron (neomycinsulfate-dexamethasone sodium phosphate); Polytrim (tfimethoprirn andpolythyxin [3 sulfate opthalmic solution); Terra-Cortril(oxytetracycline HCl and hydrocortisone acetate); Terramycin(oxytetracycline); and TobraDex (tobramycin and dexamethasone opthalmicsuspension and ointment); Vita-A opthalmic ointment, (vidatabine);(norfloxacinopthalmic solution; Ciloxan opthalmic solution and ointment(Ciprofloxacin HCl); and Ocuflox opthalmic solution (ofioxacin),Blephamide opthalmicointment (sulfacetamide sodium and prednisoloneacetate); and Blephamideopthalmic suspension (sulfacetamide sodium andpredrdsolone acetate); A/T/S (erythromycin); Bactroban (mupirocin);Benzamycin (erythromycin-benzoyl peroxide topical gel); Betadine(povidone-odine); Cleotin T (clindamy cinphosphate topical solution);Clindets (clindamycin phosphate pledgets); Cortispofin (neomycin,polymyxin B sulfates and hydrocortisone acetate cream); Emgel(erythromycin); Erycette (erythromycin topical solution); Garamycin(gentamicin sulfate); Klaron (sodium sulfacetamide lotion); Mycostatin(nystatin cream); Theramycin Z (erythromycin topical solution); T-Stat(erythromycin); Chloromycetin (chloramphenicol opthalmic ointment);Cortisporin (neomycin and polymyxin B sulfates, bacitracin zinc andhydrocortisone opthalmic ointment); Ilotycin (erythromycin); NeoDeeadron(neomycin sulfate-dexamethasone sodium phosphate); Polytrim(trimethoprim and polymyxin B sulfate); Terra-Cortril (oxytetracyclineHCl and hydrocortisone acetate); Terramycin (oxytetracyclinc); Exclderm(sulconazole nitrate); Fungizone (amphotcricin B oral suspension);Lamisil (terbinafine hydrochloride cream); Loprox (ciclopiroxolamine);Lotrimin (clotrimazole); Lotrisone (clotrimazole and betamethasonediproprionate); Mentax (butenafine HCl); Monistat-Denn (miconazolenitrate); Mycelex (clotrimazole); Mycostatin (nystatin); Naffin(nattifine HCl); Nizoral Ocetoconazole); Nystop (nystatin); Oxistat(oxiconazole nitrate); Selsun Rx (2.5% selenium sulfide lotion); andSpectazole (econazole nitrate); Denavir (penciclovir cream); and Zovirax(acyclovir); Benzashave Coenzoyl peroxide); Betadine (povidone-iodine);Betasept (chlorhexidine gluconate); Cetaphil (soap substitute);Clorpactin WCS-90 (sodium oxychlorosene); Dapsone Tablets (dapsone);Desquam-E Coenzoyl peroxide); Desquam-X (benzoyl peroxide); Hibiclens(chlorhexidine gluconate); Hibistat (ehlorhexidine gluconate); Impregon(tetrachlorosalicylanilide 2%); MetroCream (metronidazole); MetroGel(metronidazole); Noritate (metronidazole); pHisoHex (hexachlorophenedetergent cleanser); Sulfacet-R (sodium sulfacetamide 10% and sulfur5%); Sulfamylon (materfide acetate); Tfiaz Cocnzoyl peroxide); andVanoxide-HC Coenzoyl peroxide hydrocortisone); Acticin (permethrin);Elimite (permethrin); Eurax (crotamiton); Efudex (fluoro-uracil);Fluoroplex.

Pharmaceutical Compositions

Hosts, including humans can be treated by administering to the patientan effective amount of the active compound or a pharmaceuticallyacceptable prodrug or salt thereof in the presence of a pharmaceuticallyacceptable carrier or diluent. The active materials can be administeredby any appropriate route, for example, orally, parenterally,intravenously, intradermally, subcutaneously, or topically, in liquid orsolid form.

An optional dose of the compound for treatment of a bacterial (such as agram negative bacteria) infection is about 1 to 50 mg/kg, or 1 to 20mg/kg, of body weight per day, more generally 0.1 to about 100 mg perkilogram body weight of the recipient per day. The effective dosagerange of the pharmaceutically acceptable salts and prodrugs can becalculated based on the weight of the parent nucleoside to be delivered.If the salt or prodrug exhibits activity in itself, the effective dosagecan be estimated as above using the weight of the salt or prodrug, or byother means known to those skilled in the art.

Optionally, the active ingredient should be administered to achieve peakplasma concentrations of the active compound of from about 0.2 to 70 M,e.g., about 1.0 to 10 uM. This may be achieved, for example, by theintravenous injection of a 0.1 to 5% solution of the active ingredient,optionally in saline, or administered as a bolus of the activeingredient. The concentration of active compound in the drug compositionwill depend on absorption, inactivation and excretion rates of the drugas well as other factors known to those of skill in the art. It is to befurther understood that for any particular subject, specific dosageregimens should be adjusted according to the individual need and theprofessional judgment of the person administering or supervising theadministration of the compositions, and that the concentration rangesset forth herein are exemplary only and are not intended to limit thescope or practice of the claimed composition. The active ingredient maybe administered at once, or may be divided into a number of smallerdoses to be administered at varying intervals of time.

The compound is conveniently administered in unit any suitable dosageform, including but not limited to one containing 7 to 3000 mg, or 70 to1400 mg of active ingredient per unit dosage form. A dosage of 50-1000mg is optimal.

The active compound can be administered in a pharmaceutically acceptablecarrier available in the art, and can be administered by a chosen routeof administration. Pharmaceutical compositions can be prepared,packaged, or sold in a variety of formulations which can be suitable forone or more routes of administration such as, for example, oral,intravenous, intramuscular, topical, subcutaneous, rectal, vaginal,parenteral, pulmonary, intranasal, buccal, ophthalmic, or another routeof administration. The active materials can be administered in liquid orsolid form. Other contemplated formulations include projectednanoparticles, liposomal preparations, resealed erythrocytes containingthe active ingredient, and immunologically-based formulations.

The active compound may be administered intravenously orintraperitoneally by infusion or injection. Solutions of the activecompound or its salts may be prepared in water or saline, optionallymixed with a non-toxic surfactant. Dispersions may be prepared inglycerol, liquid polyethylene glycols, triacetin, and mixtures thereof,and in oils. Under ordinary conditions of storage and use, thesepreparations contain a preservative to prevent growth of microorganisms.

Pharmaceutical dosage forms suitable for injection or infusion mayinclude sterile aqueous solutions or dispersions or sterile powderscomprising the active ingredient, which are adapted for theextemporaneous preparation of sterile injectable or infusible solutionsor dispersions, optionally encapsulated in liposomes. The ultimatedosage form is optionally sterile, fluid, and stable under conditions ofmanufacture and storage. The liquid carrier or vehicle may be a solventor liquid dispersion medium comprising, for example, water, ethanol, apolyol (for example, glycerol, propylene glycol, liquid polyethyleneglycols, and the like), vegetable oils, nontoxic glyceryl esters, andsuitable mixtures thereof.

For oral therapeutic administration, the active compound can be combinedwith one or more excipients and used in the form of ingestible tablets,buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers,and the like. Such compositions and preparations can contain at least0.1% (w/w) of active compound. The percentage of the compositions andpreparations can, of course, be varied, for example from about 0.1% tonearly 100% of the weight of a given unit dosage form. The amount ofactive compound in such therapeutically useful compositions is such thatan effective dosage level will be obtained upon administration.

The tablets, troches, pills, capsules, and the like may also contain oneor more of the following: binders, such as microcrystalline cellulose,gum tragacanth, acacia, corn starch, or gelatin; excipients, such asdicalcium phosphate, starch or lactose; a disintegrating agent, such ascorn starch, potato starch, alginic acid, primogel, and the like; alubricant, such as magnesium stearate or Sterotes; a glidant, such ascolloidal silicon dixoide; a sweetening agent, such as sucrose,fructose, lactose, saccharin, or aspartame; a flavoring agent such aspeppermint, methylsalicylate, oil of wintergreen, or cherry flavoring;and a peptide antibacterial agent, such as envuvirtide (Fuzeon™). Whenthe unit dosage form is a capsule, it can contain, in addition tomaterials of the above type, a liquid carrier, such as a vegetable oilor a polyethylene glycol. Various other materials may be present ascoatings or to otherwise modify the physical form of the solid unitdosage form.

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers may be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylacetic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials may also be obtained commercially fromAlza Corporation.

The compounds/compositions of the present invention are optionallyadministered in a controlled release formulation, which can be adegradable or nondegradable polymer, hydrogel or ganogel or otherphysical construct that modifies the bioabsorption, half-life orbiodegradation of the active agent(s). The controlled releaseformulation can be a material that is painted or otherwise applied ontothe afflicted site, either internally or externally. In one embodiment,the invention provides a biodegradable bolus or implant. The controlledrelease formulation with appropriated selected imaging agent can be usedto coat a transplanted organ or tissue to prevent rejection. It canalternatively be implanted or otherwise applied near the site ofpotential infection.

Other formulations can also be developed. For example, the compounds canbe administered in liposomal suspensions (including liposomes targetedto infected cells with monoclonal antibodies to bacterial antigens).These may be prepared according to methods known to those skilled in theart, for example, as described in U.S. Pat. No. 4,522,811. For example,liposome formulations may be prepared in a variety of lipid(s) (such asstearoyl phosphatidyl ethanolaminc, stearoyl phosphatidyl choline,arachadoyl phosphatidyl choline, and cholesterol).

A pharmaceutical composition of the invention may be prepared, packaged,or sold in a formulation suitable for rectal administration. Such acomposition may be in the form of, for example, a suppository, aretention enema preparation, and a solution for rectal or colonicirrigation. A pharmaceutical composition of the invention may also beprepared, packaged, or sold in a formulation suitable for vaginaladministration. Such a composition may be in the form of, for example, asuppository, an impregnated or coated vaginally-insertable material suchas a tampon, a douche preparation, or a solution for vaginal irrigation.

A pharmaceutical composition of the invention may be prepared, packaged,or sold in a formulation suitable for pulmonary administration via thebuccal cavity. Such a formulation may comprise dry particles whichcomprise the active ingredient and which have a diameter in the rangefrom about 0.5 to about 7 nanometers, or from about 1 to about 6nanometers. Such compositions are conveniently in the form of drypowders for administration, which can include particles wherein at least98% of the particles by weight have a diameter greater than 0.5nanometers and at least 95% of the particles by number have a diameterless than 7 nanometers. Typically least 95% of the particles by weighthave a diameter greater than 1 nanometer and at least 90% of theparticles by number have a diameter less than 6 nanometers. The activeingredient can also be in the form of droplets of a solution orsuspension, for example those that have an average diameter in the rangefrom about 0.1 to about 200 nanometers.

The formulations described herein as being useful for pulmonary deliveryare also useful for intranasal delivery of a pharmaceutical compositionof the invention. Another formulation suitable for intranasaladministration is a coarse powder comprising the active ingredient andhaving an average particle from about 0.2 to 500 micrometers.

A pharmaceutical composition of the invention may be prepared, packaged,or sold in a formulation suitable for ophthalmic administration. Fortopical administration, the present compounds can be applied in pureform, i.e., as a liquid. However, typically, the compounds areadministered to the skin as compositions or formulations, in combinationwith a dermatologically acceptable carrier. Useful solid carriersinclude finely divided solids such as talc, clay, microcrystallinecellulose, silica, alumina, and the like. Useful liquid carriers includewater, alcohols, glycols, and blends of two or more of these, in whichthe present compounds can be dissolved or dispersed at effective levels,optionally with the aid of non-toxic surfactants. Adjuvants such asfragrances and additional antimicrobial agents can be added to optimizeproperties for a given use. The resulting liquid compositions can beapplied using absorbent pads, used to impregnate bandages or otherdressings, or sprayed onto the affected area using pump-type or aerosolsprayers.

Thickeners such as synthetic polymers, fatty acids, fatty acid salts andesters, fatty alcohols, modified celluloses, or modified mineralmaterials can also be employed with liquid carriers to form spreadablepastes, gels, ointments, soaps, and the like, for application directlyto the skin of the user.

The compound or a pharmaceutically acceptable prodrug or salts thereofcan also be mixed with other active materials that do not impair thedesired action, or with materials that supplement the desired action,such as antibiotics, antifungals, anti-inflammatories, or otherantibacterials, including other nucleoside compounds. Solutions orsuspensions used for parenteral, intradermal, subcutaneous, or topicalapplication can include the following components: a sterile diluent suchas water for injection, saline solution, fixed oils, polyethyleneglycols, glycerine, propylene glycol or other synthetic solvents;antibacterial agents such as benzyl alcohol or methyl parabens;antioxidants such as ascorbic acid or sodium bisulfite; chelating agentssuch as ethylenediaminetetraacetic acid; buffers such as acetates,citrates or phosphates and agents for the adjustment of tonicity such assodium chloride or dextrose. The parental preparation can be enclosed inampoules, disposale syringes or multiple dose vials made of glass orplastic. If administered intravenously, useful carriers arephysiological saline or phosphate buffered saline (PS).

The concentration of the compound(s) in a liquid composition, such as alotion, will, for example, range from about 0.1% to about 95% by weight,or from about 0.5% to about 25% by weight. The concentration in asemi-solid or solid composition such as a gel or a powder will, forexample, range from about 0.1% to 100% by weight, or about 0.5% to about5% by weight. Single doses for intravenous injection, subcutaneous,intramuscular or topical administration, infusion, ingestion orsuppository will generally be from about 0.001 to about 5000 mg, and beadministered from about 1 to about 3 times daily, to yield levels ofabout 0.01 to about 500 mg/kg, for adults.

The invention also includes one or more compounds disclosed herein, orany combination thereof, or salt thereof, in an amount effective toinhibit bacterial (such as a gram negative bacteria) replication in ahost. The compound can be useful for inhibiting bacterial replication ina cell or neutralization (i.e. inactivation) of extracellular bacteria.

The invention also includes a kit for administering a compound of theinvention, a pharmaceutically acceptable salt thereof, or apharmaceutical composition, to a host for treatment of a bacterial (suchas gram negative bacteria) infection. Typically, the host is a human.The kit comprises one or more compounds of the invention, or acombination thereof, and optionally an instructional material, whichdescribes adventitially administering the composition to the mammal byany of the routes of administration described herein. In anotherembodiment, this kit comprises a (typically sterile) solvent suitablefor dissolving or suspending the composition of the invention prior toadministering the compound to the mammal.

EXAMPLES

Nuclear magnetic resonance (NMR) spectra were obtained on a Varian INOVA400 (400 MHz) spectrometer; chemical shifts (6) are reported in partsper million (ppm), and the signals are described as s (singlet), d(doublet), t (triplet), q (quartet), bs r (broad singlet), dd (doubletof doublet), dt (triplet of doublet), and m (multiplet). All reactionswere monitored using thin layer chromatography (TLC; 200 mm silica gelGF plates) on Analtech or HPLC. Dry dichloromethane, acetonitrile, DMF,and THF were obtained by drying over 4 Å molecular sieves.

Abbreviations ACN: Acetonitrile

DBU: 1,8-Diazabicyclo[5.4.0]undec-7-ene

DCM: Dichloromethane DIEA: Diisopropylethyl-amine

DI H₂O: Deionized water

DMAP: 4-(Dimethylamino)pyridine DMF: N,N-Dimethylformamide EDC:N-(3-Dimethylaminopropyl)-N-ethylcarbodiimide HOBT:1-Hydroxybenzotriazole

IPA: iso-PropanolLDA: Lithium diisopropylamideTBS or TBDMS: tert-ButyldimethylsilylTBSOTf: tert-Butyldimethylsilyl trifluoromethanesulfonateLAH: Lithium aluminum hydridePt/C: Platinum on carbonPNB: para-Nitrobenzyl

TES: Triethylsilyl

TFA: Trifluoroacetic acid

THF: Tetrahydrofuran General Synthesis Methods Preparation of theCarbapenem Intermediate (CPI)

Carbapenem Intermediate (CPI) was prepared according to the syntheticscheme shown in Scheme 1. In the first step of the process, benzylpropionate is reacted with isobutoxycarbonyloxy acetic acid methyl esterin a solvent at low temperature in the presence of LDA to form ketoesterA. The ketoester A is then contacted with the acetoxyazetidinone B(prepared by any number of known, synthetic routes) in a solvent, andsodium carbonate is added. The reaction ages for a period of time at atemperature such that the reaction goes substantially to completion,generating the target lactam C.

The lactam C is dissolved in a solvent, such as DMF, to which a suitablebase (such as DIEA) and TBSOTf are added, and the mixture allowed to agefor a period of time at a temperature. Following workup, thebis-TBS-ketoester D is isolated.

The crude ketoester D is dissolved in ethyl acetate in an appropriatereaction vessel.

Formic acid and a catalyst, such as Pd/C, are added to the reactionvessel, and the entire mixture is hydrogenated at an appropriatehydrogen pressure (40-50 psi) for a period of time such that thedecarboxylation reaction proceeds to completion. The reaction mixture isfiltered over a pad of Celite, and the solvent is removed under vacuum.Product E is isolated following purification by column chromatography.

The bis-TBDMS ketolactam E is then de-silylated using 2 N HCl in ACN andthe product is isolated after a standard aqueous workup. The crudeproduct is dissolved in a solvent, such as DCM, and allowed to reactwith triethylsilyl chloride and imidazole for several hours (monitoredby TLC) at rt. Following aqueous workup, O-TES ketolactam F was isolatedand purified on silica gel.

N-PNB, O-TES ketolactam G is produced by reacting ketolactam F withp-nitrobenzyl oxalylchloride in a suitable solvent (DCM, for example) inthe presence of a base (DIEA, for example). The mixture is allowed toage for a period of time (and at an appropriate temperature) to effect asubstantially complete reaction as monitored by an appropriate means(e.g., TLC or HPLC). Following workup in a usual manner, intermediate Gwas isolated.

To a solution of compound G is a suitable solvent was addedtriethylphosphite, and the mixture heated to reflux until complete byTLC. Following workup and purification in the appropriate manner, CPIwas isolated.

Preparation of Gram-Negative Active Carbapenems

The 1-β-methylcarbapenem compounds possessing Gram-negative activitywere synthesized using the methods described above and as illustrated inScheme 2 below, unless otherwise noted. In general, a series ofsecondary or cyclic amines (H) were coupled to CPI in DMF using acombination of Pd₂(dba)₃CHCl₃ with P(OEt)₃ at rt to produce the coupledintermediate I. In some cases, 2,6-lutidine (Method B), TsOH (Method C),or DIEA (Method D) were added to drive the reaction to completion. Thesecondary or cyclic amines were either purchased from commercial sourcesor prepared by alkylation or substitution reactions of N-Boc-protectedprimary amines followed by cleavage of the Boc protecting group withTFA/water in DCM

Removal of the TES protecting group in the series of intermediate I wasaccomplished as described in Method E.

Lastly, the PNB group(s) in intermediate J were removed by hydrogenationof the corresponding PNB esters using Methods F, G or H and the finalproducts K were isolated.

Step 1: General Procedure for the Palladium Coupling Reaction Method A:

To an oven-dried round-bottomed flask was added anhydrous DMF. This wasdegassed at rt with two cycles of nitrogen/vacuum. Then Pd₂(dba)₃CHCl₃and P(OEt)₃ were added. The solution was degassed with twonitrogen/vacuum cycles and aged for 20 min. Then neutralized aminedissolved in DMF and CPI were added and the resulting mixture wasdegassed with two nitrogen/vacuum cycles, and was allowed to stir at rt.After the consumption of CPI, solvent was removed in vacuo and theresulting residue was purified by SiO₂ column chromatography to providethe desired coupled product.

Method B:

To an oven-dried round-bottomed flask was added anhydrous DMF. This wasdegassed at rt with two cycles of nitrogen/vacuum. Then Pd₂(dba)₃CHCl₃and P(OEt)₃ were added. The solution was degassed with twonitrogen/vacuum cycles and aged for 20 min. Then amine (TFA salt) andCPI were added, followed by 2,6-lutidine and the resulting mixture wasdegassed with two nitrogen/vacuum cycles, and was allowed to stir at rt.After the consumption of CPI, solvent was removed in vacuo and theresulting residue was purified by SiO₂ column chromatography to providethe desired coupled product and its de-TES product.

Method C:

To an oven-dried round-bottomed flask were added anhydrous DMF and 4 Åmolecular sieves. This was degassed at rt with two cycles ofnitrogen/vacuum. Then Pd₂(dba)₃CHCl₃ and P(OEt)₃ were added. Thesolution was degassed with two nitrogen/vacuum cycles and aged for 20min. Then amine and CPI were added, followed by TsOH and the resultingmixture was degassed with two nitrogen/vacuum cycles, and was allowed tostir at rt. After the consumption of CPI, solvent was removed in vacuoand the resulting residue was purified by SiO₂ column chromatography toprovide the desired coupled product.

Method D:

To an oven-dried round-bottomed flask was added anhydrous toluene andTHF (10 to 1 ratio). This was degassed at ice-bath with two cycles ofnitrogen/vacuum. Then Pd₂(dba)₃CHCl₃ and P(OEt)₃ were added. Thesolution was degassed with two nitrogen/vacuum cycles and aged for 20min. Then amine (TFA salt) and CPI were added, followed by DIEA and theresulting mixture was degassed with two nitrogen/vacuum cycles, and wasallowed to stir at rt. After the consumption of CPI, solvent was removedin vacuo and the resulting residue was purified by SiO₂ columnchromatography to provide the desired coupled product.

Step 2: General Procedure for the Removal of the TES Protecting GroupMethod E:

To a round-bottomed flask charged with TES compound was added anhydrousTHF and DMF under N₂. This was cooled to 0° C. and then AcOH was addedfollowed by Me₄NF.4H₂O. After stirring overnight at 0° C., the crudemixture was quenched with DI water, followed by addition of saturatedNaHCO₃ to adjust pH 7. Then this was extracted with EtOAc or a mixtureof DCM and MeOH. The combined organic layer was dried (Na₂SO₄) andconcentrated in vacuo. The crude material was purified by SiO₂ columnchromatography to provide the desired OH product.

Step 3: General Procedure for the Removal of the PNB Protecting GroupMethod F:

To a round-bottomed flask charged with OH compound was added THF, IPA,DI water and phosphate buffer (pH 6, 0.35 M). This was degassed andcharged with N₂. Then Pt/C was added, followed by degassing and chargingwith H₂ (H₂ balloon). After stirring at 0° C. until the consumption ofSM, cold DI water was added. The crude mixture was filtered throughCelite and the filtrate was extracted with cold EtOAc twice. Theseparated aqueous layer was concentrated in vacuo. The crude materialwas purified by SP-207 resin with IPA and DI water as eluent. The columnfractions were concentrated under reduced pressure at 6° C. to removeIPA and then lyophilized to afford the final product.

Method G:

To solution of OH compound in THF and phosphate buffer (pH 6.0, 0.35 M)was added Zinc dust at 10° C. and aged over until the consumption of SM.The mixture was diluted with cold DI water, filtered through on a pad ofCelite, and the pad was washed with water and ethyl acetate. Afterseparation, the aqueous layer was lyophilized and then purified on HP-20or SP-207 resins with a solvent gradient system (from 100% water to 45%i-PrOH in water). The column fractions containing product were thenconcentrated under vacuum and lyophilized to afford the final product.

Method H:

It was dissolved OH compound in a glass vessel of parr-hydrogenationwith a mixed solvent of THF/iso-propanol/DI-water/phosphate buffer (pH6, 0.35 M). To the mixture was added Pt catalyst (5% or 10% on Carbon),degassed under vacuum, and charged with H₂ gas to 30 psi. After shakingabout 30 min until no more pressure change, the reaction mixture wascooled down to zero degree and diluted with DI water. The mixture wasfiltered through on a pad of Celite and the pad was washed with water.After washing with ethyl acetate, the aqueous layer was lyophilized andthen purified on SP-207 resin with a solvent gradient system (from 100%water to 45% i-PrOH in water). The column fractions containing productwere then concentrated under vacuum and lyophilized to afford desiredfinal carbapenem derivative.

Example 1 Synthesis of Compound 7

Step 1

In a 500 mL oven-dried round-bottomed flask charged with(R)-3-pyrrolidinol (5.15 g, 58 mmol) was added dry CH₃CN (200 mL) togive a light brown solution under N₂. Then Et₃N (16.2 mL, 0.12 mol) wasadded dropwise. This was cooled to 0° C. and then CbzCl (48 mL, 83 mmol)was added dropwise. The temperature was allowed to warm up to rtgradually. After stirring for 24 h, solvent was removed in vacuo. Theresidue was treated with DCM and DI water. Organic layer was separatedand the aqueous layer was extracted with DCM once. The combined organiclayer was dried (Na₂SO₄) and concentrated in vacuo. The crude materialwas purified by SiO₂ column chromatography eluting with Hexane:EtOAc=1:1to provide the desired alcohol 1 (12.6 g, 98%).

¹H NMR (CDCl₃, 400 MHz): δ 7.37-7.30 (m, 5H), 5.14 (s, 2H), 4.52-4.46(m, 1H), 3.60-3.41 (m, 4H), 2.01-1.95 (m, 2H), 1.61 (br s, 1H).

Step 2

In a 1 L oven-dried round-bottomed flask charged with alcohol 1 (12.6 g,56.9 mmol) was added dry DCM (250 mL) to give a colorless solution underN₂. This was cooled to 0° C. and Et₃N (16 mL, 0.11 mol) was added. After10 min, MsCl (6.5 mL, 84 mmol) was added dropwise. The temperature wasallowed to warm up to rt gradually. After 14 h, DI water was added. Theorganic layer was separated, washed with DI water once, brine once,dried (Na₂SO₄) and concentrated in vacuo. The crude material waspurified by SiO₂ column chromatography eluting from Hexane:EtOAc=2:1 to1:1 to provide the desired mesylate 2 (14.7 g, 86%) as a yellow oil.

¹H NMR (CDCl₃, 400 MHz): δ 7.38-7.30 (m, 5H), 5.30-5.28 (m, 1H), 5.14(d, J=2.8 Hz, 2H), 3.81-3.74 (m, 1H), 3.70-3.51 (m, 3H), 3.04 (s, 3H),2.36-2.27 (m, 1H), 2.21-2.09 (m, 1H).

Step 3

In a 150 mL oven-dried round-bottomed flask charged with mesylate 2(4.12 g, 0.014 mol) was added dry DMSO (30 mL) under N₂. This wastreated with KCN (1.94 g, 0.03 mol) at rt. The mixture was heated to 80°C. After 21 h, the flask was removed from oil-bath. After cooling to rt,sat NaHCO₃ was added, which was extracted with EtOAc (×4). The combinedorganic layer was dried (Na₂SO₄) and concentrated in vacuo. The crudematerial was purified by SiO₂ column chromatography eluting fromHexane:EtOAc=3:1 to 2:1 to provide the desired cyanate 3 (2.23 g, 70%)as a yellow oil.

¹H NMR (CDCl₃, 400 MHz): δ 7.37-7.30 (m, 5H), 5.14 (d, J=1.6 Hz, 2H),3.78-3.47 (m, 4H), 3.15-3.07 (m, 1H), 2.27-2.16 (m, 2H).

Step 4

To a 250 mL two-neck flask was added a MeOH solution of cyanate 3 (0.53g, 2.3 mmol) under N₂. Then 20 mol % Pd(OH)₂/C (0.16 g, 0.23 mmol) wasadded, followed by vacuum and charging with H₂, this was repeated twice.After stirring for 1 h under hydrogen balloon, TLC showed no SM. Thenthe crude mixture was filtered through Celite and washed with MeOH. Thefiltrate was concentrated to give the crude amine 4 (0.13 g, 61%) as alight yellow oil which was used directly for the next step.

¹H NMR (CDCl₃, 400 MHz): δ 3.25-3.09 (m, 2H), 3.00-2.86 (m, 2H),2.70-2.38 (m, 1H), 2.26-2.02 (m, 2H).

Step 5

According to General Method A, CPI (1.54 g, 2.6 mmol), side chain 4(0.25 g, 2.6 mmol), Pd₂(dba)₃CHCl₃ (0.14 g, 0.135 mmol) and P(OEt)₃(0.15 mL, 0.86 mmol) in DMF (53 mL) were reacted for 17.5 h to affordthe TES product 5 (0.39 g, 26%).

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (d, J=8.8 Hz, 2H), 7.66 (d, J=9.2 Hz,2H), 5.44 (d, J=13.6 Hz, 1H), 5.22 (d, J=14.0 Hz, 1H), 4.29-4.20 (m,2H), 4.14-4.07 (m, 1H), 3.89 (d, J=14.4 Hz, 1H), 3.38 (d, J=14.8 Hz,1H), 3.36-3.29 (m, 1H), 3.26-3.23 (m, 1H), 3.05-2.98 (m, 1H), 2.87-2.83(m, 1H), 2.77-2.71 (m, 2H), 2.62-2.56 (m, 1H), 2.27-2.18 (m, 1H),2.17-2.08 (m, 1H), 1.25 (d, J=7.0 Hz, 3H), 1.17 (d, J=7.2 Hz, 3H), 0.94(t, J=8.0 Hz, 9H), 0.60 (q, J=8.0 Hz, 6H).

Step 6

According to General Method E, TES compound 5 (390 mg), Me₄NF.4H₂O (0.17g), AcOH (79 μL) in THF (15 mL) and DMF (5 mL) were reacted for 7.5 h toafford the desired OH product 6 (240 mg, 77%) as a white glassy solid.

¹H NMR (CDCl₃, 400 MHz): δ 8.24 (d, J=8.8 Hz, 2H), 7.67 (d, J=8.8 Hz,2H), 5.49 (d, J=13.6 Hz, 1H), 5.22 (d, J=13.6 Hz, 1H), 4.30-4.26 (m,1H), 4.24 (dd, J=10.0, 3.2 Hz, 1H), 3.89 (d, J=14.8 Hz, 1H), 3.40 (d,J=14.8 Hz, 1H), 3.40-3.37 (m, 1H), 3.30-3.28 (m, 1H), 3.05-2.98 (m, 1H),2.87-2.83 (m, 1H), 2.78-2.69 (m, 2H), 2.61-2.55 (m, 1H), 2.27-2.09 (m,2H), 1.72 (d, J=4.4 Hz, 1H), 1.36 (d, J=6.4 Hz, 3H), 1.19 (d, J=7.2 Hz,3H).

Step 7

According to General Method F, OH compound 6 (240 mg, 0.53 mmol), 10%Pt/C (280 mg) in IPA (10 mL), THF (20 mL), DI water (16 mL) and pH 6buffer (7 mL) were reacted for 8 h to afford the desired final product 7(18 mg, 11%).

¹H NMR (D₂O, 400 MHz): % 4.21-4.13 (m, 2H), 3.72 (d, J=13.6 Hz, 1H),3.45 (d, J=13.2 Hz, 1H), 3.37 (dd, J=6.4, 3.8 Hz, 1H), 3.26-3.14 (m,2H), 2.99 (br s, 1H), 2.86 (br s, 2H), 2.67 (br s, 1H), 2.34-2.23 (m,1H), 2.16-2.07 (m, 1H), 1.23 (d, J=6.4 Hz, 3H), 1.06 (d, J=7.2 Hz, 3H).

Example 2 Synthesis of Compound 12

Step 1

In a 150 mL round-bottomed flask charged with cyanate 3 (0.64 g, 2.78mmol) was added acetone (15 mL), followed by DI water (4.9 mL) to give acolorless solution. Then 30% aqueous H₂O₂ (7.8 mL) was added, followedby Na₂CO₃ (0.97 g, 9.15 mmol). After stirring at rt for 20 h, the crudemixture was treated with EtOAc and brine. Organic layer was separatedand the aqueous layer was extracted with EtOAc twice. The combinedorganic layer was dried (Na₂SO₄) and concentrated in vacuo. The crudematerial was purified by SiO₂ column chromatography eluting fromHexane:EtOAc=1:1 to EtOAc to provide the desired amide 8 (0.38 g, 55%)as a white solid.

¹H NMR (DMSO-d6, 400 MHz): δ 7.46 (br s, 1H), 7.39-7.31 (m, 5H), 6.97(br s, 1H), 5.05 (s, 2H), 3.53-3.24 (m, 4H), 2.94-2.87 (m, 1H),2.04-1.93 (m, 2H).

Step 2

To a 100 mL two-neck flask was added an EtOH solution of amide 8 (0.38g, 1.53 mmol). This was vacuumed and charged with N₂. Then 10 mol % Pd/C(82 mg) was added, followed by vacuum and charging with H₂ (hydrogenballoon), this was repeated twice. After stirring for 1.5 h, TLC showeda lot of SM. The crude mixture was transferred to a Parr hydrogenationflask and was hydrogenated at 50 psi. After 1 h, TLC showed no SM. Thenthe crude mixture was filtered through Celite and washed with EtOH. Thefiltrate was concentrated in vacuo to give the crude amine 9 (0.15 g,84%) which was used directly for the next step.

¹H NMR (DMSO-d6, 400 MHz): δ 7.31 (br s, 1H), 6.76 (br s, 1H), 2.92-2.87(m, 1H), 2.80-2.59 (m, 4H), 1.82-1.67 (m, 2H).

Step 3

According to General Method A, CPI (0.78 g, 1.3 mmol), side chain 9(0.15 g, 1.3 mmol), Pd₂(dba)₃CHCl₃ (69 mg, 0.067 mmol) and P(OEt)₃ (77μL, 0.44 mmol) in DMF (27 mL) were reacted for 27 h to afford thedesired TES product 10 (0.38 g, 49%) as a yellow oil.

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (d, J=8.8 Hz, 2H), 7.66 (d, J=9.2 Hz,2H), 6.40 (br s, 1H), 5.44 (d, J=14.0 Hz, 1H), 5.35 (br s, 1H), 5.22 (d,J=14.0 Hz, 1H), 4.27-4.20 (m, 2H), 3.91 (d, J=14.4 Hz, 1H), 3.36 (d,J=14.8 Hz, 1H), 3.30-3.23 (m, 2H), 2.90-2.83 (m, 3H), 2.50-2.40 (m, 2H),2.22-2.13 (m, 1H), 2.07-1.98 (m, 1H), 1.25 (d, J=6.0 Hz, 3H), 1.18 (d,J=7.2 Hz, 3H), 0.94 (t, J=8.0 Hz, 9H), 0.60 (q, J=8.0 Hz, 6H).

Step 4

According to General Method E, TES compound 10 (380 mg, 0.65 mmol),Me₄NF.4H₂O (0.16 g, 0.99 mmol), AcOH (74 μL, 1.29 mmol) in THF (14 mL)and DMF (4.5 mL) were reacted for 15 h to afford the desired OH product11 (300 mg, 95%) as a white glassy solid.

¹H NMR (CDCl₃, 400 MHz): δ 8.23 (d, J=8.8 Hz, 2H), 7.66 (d, J=8.8 Hz,2H), 6.35 (br s, 1H), 5.48 (d, J=13.6 Hz, 1H), 5.32 (br s, 1H), 5.21 (d,J=13.6 Hz, 1H), 4.30-4.22 (m, 2H), 3.91 (d, J=14.4 Hz, 1H), 3.37 (d,J=14.8 Hz, 1H), 3.34-3.27 (m, 2H), 2.95-2.82 (m, 3H), 2.49-2.39 (m, 2H),2.22-2.13 (m, 1H), 2.07-1.98 (m, 1H), 1.36 (d, J=6.4 Hz, 3H), 1.19 (d,J=7.6 Hz, 3H).

Step 5

According to General Method F, OH compound 11 (0.29 g, 0.62 mmol), 10%Pt/C (300 mg) in IPA (7.5 mL), THF (15 mL), DI water (15 mL) and pH 6buffer (6 mL) were reacted for 8 h to afford the desired product 12 (70mg, 34%).

¹H NMR (D₂O, 400 MHz): δ 4.22-4.16 (m, 2H), 4.00 (br s, 2H), 3.44-3.42(m, 2H), 3.28-3.14 (m, 4H), 2.36 (br s, 1H), 2.16 (br s, 1H), 1.22 (d,J=6.0 Hz, 3H), 1.11 (d, J=7.2 Hz, 3H).

Example 3 Synthesis of Compound 19

Step 1

To a 100 mL round-bottomed flask charged with cyanate 3 (2 g, 8.7 mmol)was added concentrated HCl (20 mL). After refluxing for 4.5 h, solventwas removed in vacuo and was dried overnight by oil-pump. The crudematerial was re-dissolved in a mixture of acetone (20 mL) and DI water(20 mL). After cooling to 0° C., Na₂CO₃ (2.8 g, 26 mmol) was added,followed by dropwise addition of CbzCl (5.5 mL, 9.6 mmol). The reactionwas allowed to warm up to rt gradually. After 7 h, solvent was removedin vacuo. Then DI water (8 mL) was added, which was extracted withHexane:EtOAc=1:1 twice. The aqueous layer was acidified to pH 2 byadding concentrated HCl and 0.5 M KHSO₄. The aqueous layer was extractedwith EtOAc (×4), dried (Na₂SO₄) and concentrated in vacuo to give thecarboxylic acid 13 (1.46 g, 67%) which was used directly for the nextstep.

¹H NMR (CDCl₃, 400 MHz): δ 7.38-7.29 (m, 5H), 5.14 (d, J=2.8 Hz, 2H),3.72-3.43 (m, 4H), 3.17-3.09 (m, 1H), 2.21-2.14 (m, 2H).

Step 2

In a 150 mL round-bottomed flask charged with carboxylic acid 13 (1.48g, 5.94 mmol) was added DCM (30 mL) to give a colorless solution underN₂. This was cooled to 0° C. and then N-hydroxysuccinamide (1.0 g, 8.7mmol) and EDC.HCl (1.37 g, 7.15 mmol) were added.

This was allowed to warm up to rt gradually. After stirring for 18 h,solvent was removed in vacuo. The residue was treated with EtOAc and DIwater. Organic layer was separated and washed with DI water (×3) andbrine once, dried (MgSO₄) and concentrated in vacuo to give the desiredproduct 14 (1.91 g, 93%) which was used directly for the next step.

¹H NMR (CDCl₃, 400 MHz): δ 7.37-7.29 (m, 5H), 5.14 (d, J=7.6 Hz, 2H),3.82-3.49 (m, 4H), 3.44-3.37 (m, 1H), 2.86-2.82 (m, 4H), 2.37-2.30 (m,2H).

Step 3

In a 75 mL sealed tube was added a DMF (11 mL) solution of startingmaterial 14 (1.9 g, 5.5 mmol), followed by sulfamide (1.07 g, 11 mmol).The mixture was heated to 90° C. for 16.5 h, after cooling to rt, thecrude mixture was filtered and the solid was washed with DCM. Thefiltrate was concentrated in vacuo. The residue was treated with DCM andDI water. Aqueous layer was separated and extracted with DCM (×3). Thecombined organic layer was washed with brine once, dried (Na₂SO₄) andconcentrated in vacuo. The crude material was purified by SiO₂ columnchromatography eluting from 3% MeOH in DCM to 5% to 9% to provide thedesired product 15 (0.74 g, 41%).

¹H NMR (CDCl₃, 400 MHz): δ 7.34-7.28 (m, 5H), 6.13 (br s, 1H), 6.00 (brs, 1H), 5.10 (d, J=2.0 Hz, 2H), 3.70-3.52 (m, 3H), 3.42-3.36 (m, 1H),2.97-2.85 (m, 1H), 2.20-2.03 (m, 2H).

Step 4

To a Parr hydrogenation flask was added a MeOH (23 mL) solution ofstarting material 15 (0.74 g, 2.26 mmol). This was vacuumed and chargedwith H₂. Then 0.24 g of Pd/C (10% on carbon) was added. This washydrogenated at 50 psi for 2 h. Then the crude mixture was filteredthrough Celite and washed with MeOH. The filtrate was concentrated invacuo to give the crude amine 16 (0.21 g, 48%) as a colorless oil whichwas used directly for the next step.

¹H NMR (CDCl₃, 400 MHz): δ 6.00 (br s, 1H), 5.19 (br s, 1H), 3.21-3.10(m, 2H), 2.98-2.93 (m, 1H), 2.87-2.74 (m, 2H), 2.11-2.02 (m, 1H),2.00-1.91 (m, 1H).

Step 5

According to General Method C, CPI (0.59 g, 1.0 mmol), side chain 16(0.21 g, 1.1 mmol), Pd₂(dba)₃CHCl₃ (52 mg, 0.05 mmol), P(OEt)₃ (59 μL,0.33 mmol) and TsOH (97 mg, 0.5 mmol) in DMF (20 mL) were reacted for 16h to afford the TES product 17 (0.53 g, 79%) as a light yellow glassysolid.

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (d, J=8.8 Hz, 2H), 7.67 (d, J=8.8 Hz,2H), 6.39 (bs, 1H), 5.45 (d, J=13.6 Hz, 1H), 5.22 (d, J=14.0 Hz, 1H),5.13 (br s, 1H), 4.27-4.20 (m, 2H), 3.92 (d, J=14.4 Hz, 1H), 3.37 (d,J=14.0 Hz, 1H), 3.30-3.23 (m, 2H), 2.97-2.85 (m, 3H), 2.50-2.40 (m, 2H),2.23-2.17 (m, 1H), 2.05-2.01 (m, 1H), 1.26 (d, J=6.0 Hz, 3H), 1.18 (d,J=7.6 Hz, 3H), 0.94 (t, J=8.0 Hz, 9H), 0.60 (q, J=8.0 Hz, 6H).

Step 6

According to General Method E, TES compound 17 (0.52 g, 0.78 mmol),Me₄NF.4H₂O (0.19 g, 1.17 mmol), AcOH (89 μL, 1.55 mmol) in THF (17 mL)and DMF (5.6 mL) were reacted for 16.5 h to afford the desired OHproduct 18 (0.31 g, 72%) as an off-white glassy solid.

¹H NMR (CDCl₃, 400 MHz): δ 8.23 (d, J=8.8 Hz, 2H), 7.67 (d, J=8.8 Hz,2H), 6.30 (br s, 1H), 5.49 (d, J=13.6 Hz, 1H), 5.22 (d, J=13.6 Hz, 1H),5.14 (br s, 1H), 4.29-4.22 (m, 2H), 3.91 (d, J=14.4 Hz, 1H), 3.38 (d,J=14.8 Hz, 1H), 3.34-3.27 (m, 2H), 2.96-2.83 (m, 3H), 2.52-2.40 (m, 2H),2.21-2.13 (m, 1H), 2.09-2.01 (m, 1H), 1.69 (bs, 1H), 1.36 (d, J=6.0 Hz,3H), 1.20 (d, J=7.6 Hz, 3H).

Step 7

According to General Method F, OH compound 18 (0.31 g, 0.56 mmol), 10%Pt/C (300 mg) in IPA (8 mL), THF (20 mL), DI water (22 mL) and pH=6buffer (7 mL) were reacted for 5 h to afford the desired product 19 (95mg, 41%).

¹H NMR (D₂O, 400 MHz): δ 4.26-4.22 (m, 2H), 4.15 (d, J=15.2 Hz, 1H),4.09 (d, J=15.2 Hz, 1H), 3.50-3.38 (m, 7H), 3.28-3.20 (m, 1H), 2.50-2.42(m, 1H), 2.27-2.18 (m, 1H), 1.28 (d, J=6.0 Hz, 3H), 1.18 (d, J=7.2 Hz,3H).

Example 4 Synthesis of Compound 27

Step 1

(R)-Pyrrolidinol (43.56 g, 0.5 mol) was dissolved in dry CH₂Cl₂ (1 L)and cooled with ice-bath to 0° C. To the solution was added Et₃N (139.4mL, 1.0 mol), followed by dropwise addition of (Boc)₂O (130.95 g, 0.6mol) in CH₂Cl₂ (160 mL), and keep stirring at 0° C. for 2 h.

To the reaction mixture of Boc-protection was added more of Et₃N (139.4mL, 1.0 mol), and followed by dropwise addition of MsCl (42.74 mL, 0.55mol). After 2 h at 0° C., it was treated with H₂O (500 mL) for 10 min,separated, and the aqueous phase was extracted with CH₂Cl₂ (300 mL×2).The combined organic layers was washed with brine (500 mL), concentratedand purified by silica column chromatography to give N-Boc protectedmesylate 20 as an oily product (123 g, 93%)

¹H NMR (CDCl₃, 400 MHz): δ 5.29-5.23 (m, 1H), 3.74-3.40 (m, 4H), 3.05(s, 3H), 2.36-2.20 (m, 1H), 2.20-2.05 (m, 1H), 1.46 (s, 9H).

Step 2

To a solution of mesylate 20 (19.8 g, 74.7 mmol) in DMF (250 mL) wasadded NaN₃ (7.28 g, 112 mmol), and aged at 95° C. for 20 h. Aftercooling down to rt, the mixture was concentrated under a reducedpressure, treated with H₂O (200 mL), and extracted with CH₂Cl₂ (100mL×3). The combined organic layers was washed with brine (100 mL),concentrated and purified by silica column chromatography to afford thedesired azide 21 (14.4 g, 90%).

¹H NMR (CDCl₃, 400 MHz): δ 4.16-4.10 (m, 1H), 3.54-3.32 (m, 4H),2.12-1.94 (m, 2H), 1.45 (s, 9H).

Step 3

A solution of azide 21 (14.4 g, 68.1 mmol) and H₂O (7.4 mL, 0.41 mol) inTHF (200 mL) was cooled with ice-bath, then PPh₃ (35.73 g, 136.2 mmol)was added into the mixture as a solid in small portions. After theaddition, the reaction mixture was slowly warmed up to rt and thensubmerged into an oil-bath preheated to 50° C. After 5 h, the mixturewas concentrated in vacuo, treated with H₂O (100 mL) and CH₂Cl₂ (100mL), and then acidified with 1N HCl to pH 2. The mixture was washed withDCM (100 mL×3), and the aqueous phase was then treated with 6 N NaOH topH 10. After extraction with CH₂Cl₂ (100 mL×3), the organic layers werecombined and washed with brine (100 mL), dried over Na₂SO₄, concentratedto give the desired amine 22 (12.16 g, 96%) which was used directly forthe next step.

¹H NMR (CDCl₃, 400 MHz): δ 3.58-3.28 (m, 4H), 3.08-2.94 (m, 1H),2.08-1.98 (m, 1H), 1.70-1.56 (m, 1H), 1.45 (s, 9H).

Step 4

A solution of amine 22 (372 mg, 2 mmol) in dry CH₂Cl₂ (20 mL) was cooledto 0° C., then Et₃N (558 μL, 4 mmol) and ClCO₂PNB (517 mg, 2.4 mmol)were added into the solution. The reaction mixture was aged at 0° C. for2 h, then it was treated with H₂O (20 mL) and separated, the aqueouslayer was extracted with CH₂Cl₂ (20 mL×2). The combined organic layerswas washed with brine (30 mL), concentrated and purified by silica gelcolumn chromatography to give the desired carbamate 23 (0.35 g, 48%).

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (d, J=8.8 Hz, 2H), 7.51 (d, J=8.8 Hz,2H), 5.20 (s, 2H), 4.93 (d, J=6.4 Hz, 1H), 4.30-4.18 (m, 1H), 3.61 (dd,J=6.4, 11.6 Hz, 1H), 3.50-3.35 (m, 2H), 3.32-3.13 (m, 1H), 2.21-2.09 (m,1H), 1.93-1.78 (m, 1H), 1.46 (s, 9H).

Step 5

To a solution of TFA (1.1 mL, 14.4 mmol) in CH₂Cl₂ (10 mL) at 0° C. wasadded the carbamate 23 (0.35 g, 0.96 mmol). After overnight at 0° C.,the mixture was concentrated, co-evaporated with hexane (5 mL×3) andthen dried under high vacuum to give the de-Boc product as a TFA salt.The TFA salt was then neutralized with saturated NaHCO₃, extracted with5% methanol in DCM to give compound 24 (0.21 g, 85%).

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (d, J=8.4 Hz, 2H), 7.50 (d, J=8.4 Hz,2H), 5.23 (s, 1H), 5.17 (s, 2H), 4.23-4.12 (m, 1H), 3.16-3.02 (m, 2H),2.98-2.76 (m, 2H), 2.20-2.08 (m, 1H), 1.72-1.56 (m, 1H).

Step 6

According to General Method A, CPI (0.25 g, 0.42 mmol), side chain 24(0.11 g, 0.41 mmol), Pd₂(dba)₃CHCl₃ (22 mg, 0.021 mmol) and P(OEt)₃ (24μL, 0.14 mmol) in DMF (9 mL) were reacted for 4 h to afford the desiredTES product 25 (0.26 g, 85%) as a yellow oil.

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (d, J=8.8 Hz, 4H), 7.66 (d, J=8.4 Hz,2H), 7.50 (d, J=8.4 Hz, 2H), 5.44 (d, J=14.4 Hz, 1H), 5.23-5.17 (m, 3H),5.02 (d, J=8.4 Hz, 1H), 4.27-4.18 (m, 3H), 3.85 (d, J=14.4 Hz, 1H), 3.34(d, J=14.4 Hz, 1H), 3.31-3.23 (m, 2H), 2.79-2.74 (m, 1H), 2.64-2.60 (m,1H), 2.54-2.51 (m, 1H), 2.48-2.42 (m, 1H), 2.32-2.21 (m, 1H), 1.25 (d,J=6.0 Hz, 3H), 1.17 (d, J=7.2 Hz, 3H), 0.94 (t, J=8.0 Hz, 9H), 0.60 (q,J=7.6 Hz, 6H).

Step 7

According to General Method E, TES compound 25 (0.28 g, 0.38 mmol),Me₄NF.4H₂O (94 mg, 0.58 mmol), AcOH (43 μL, 0.75 mmol) in THF (8 mL) andDMF (2.7 mL) were reacted for 16 h to afford the desired OH product 26(0.17 g, 71%).

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (dd, J=8.8, 2.0 Hz, 4H), 7.66 (d, J=8.8Hz, 2H), 7.50 (d, J=8.4 Hz, 2H), 5.48 (d, J=13.6 Hz, 1H), 5.21 (d,J=14.0 Hz, 1H), 5.18 (s, 2H), 5.02 (d, J=8.4 Hz, 1H), 4.27 (t, J=6.0 Hz,1H), 4.22-4.19 (m, 2H), 3.84 (d, J=14.4 Hz, 1H), 3.36 (d, J=14.8 Hz,1H), 3.36-3.26 (m, 2H), 2.82-2.77 (m, 1H), 2.58-2.52 (m, 2H), 2.45-2.39(m, 1H), 2.32-2.21 (m, 1H), 1.36 (d, J=6.0 Hz, 3H), 1.18 (d, J=7.2 Hz,3H).

Step 8

According to General Method F, OH compound 26 (0.17 g, 0.27 mmol), 5%Pt/C (370 mg) in IPA (4 mL), THF (8 mL), DI water (8 mL) and phosphatebuffer (pH 6, 3, mL) were reacted for 8 h to afford the desired finalproduct 27 (12 mg, 14%).

¹H NMR (D₂O, 400 MHz): δ 4.27-4.22 (m, 2H), 3.84-3.80 (m, 2H), 3.63-3.60(m, 1H), 3.45-3.43 (m, 1H), 3.26-3.18 (m, 2H), 2.91 (br s, 2H), 2.76 (brs, 1H), 2.39-2.30 (m, 1H), 1.89-1.84 (m, 1H), 1.28 (d, J=6.0 Hz, 3H),1.13 (d, J=7.2 Hz, 3H).

Example 5 Synthesis of Compound 32

Step 1

A solution of amine 22 (930 mg, 5 mmol) in dry CH₂Cl₂ (50 mL) was cooledto 0° C., then Et₃N (1.4 mL, 10 mmol) and acetic anhydride (567 μL, 6mmol) were added into the solution, slowly warmed up to rt. Afterovernight, it was treated with H₂O (20 mL) and separated, the aqueouslayer was extracted with CH₂Cl₂ twice (10 mL). The combined organiclayers was washed with brine (30 mL), then concentrated and purified bysilica gel column chromatography to give a desired amide 28 (0.69 g,60%). ¹H NMR (CDCl₃, 400 MHz): δ 6.36-6.06 (br s, 1H), 4.45-4.36 (m,1H), 3.68-3.48 (m, 1H), 3.43-3.30 (m, 2H), 3.24-3.08 (m, 1H), 2.14-2.04(m, 1H), 1.95 (s, 3H), 1.90-1.73 (m, 1H), 1.42 (s, 9H).

Step 2

The similar procedure with side chain, 24, synthesis was used forde-protection of Boc group to afford the desired amine 29 in 86% yield.

¹H NMR (CDCl₃, 400 MHz): δ 4.54-4.46 (m, 1H), 3.48-3.38 (m, 1H),3.32-3.17 (m, 3H), 2.25-2.14 (m, 1H), 2.06-1.96 (m, 1H), 1.92 (s, 3H).

Step 3

According to General Method A, CPI (0.74 g, 1.25 mmol), side chain 29(0.16 g, 1.25 mmol), Pd₂(dba)₃CHCl₃ (65 mg, 0.063 mmol) and P(OEt)₃ (72μL, 0.41 mmol) in DMF (25 mL) were reacted for 24.5 h to afford thedesired TES product 30 (0.27 g, 36%).

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (dd, J=8.8, 1.6 Hz, 2H), 7.66 (d, J=8.8Hz, 2H), 5.64 (d, J=8.0 Hz, 1H), 5.45 (d, J=14.0 Hz, 1H), 5.22 (d,J=14.0 Hz, 1H), 4.46-4.39 (m, 1H), 4.28-4.22 (m, 1H), 4.19 (dd, J=10.4,3.2 Hz, 1H), 3.84 (d, J=14.4 Hz, 1H), 3.35 (d, J=14.4 Hz, 1H), 3.30-3.22(m, 2H), 2.80-2.74 (m, 1H), 2.62-2.58 (m, 1H), 2.52-2.48 (m, 1H),2.47-2.40 (m, 1H), 2.31-2.23 (m, 1H), 1.96 (s, 3H), 1.61-1.53 (m, 1H),1.26 (d, J=6.4 Hz, 3H), 1.18 (d, J=7.2 Hz, 3H), 0.94 (t, J=8.0 Hz, 9H),0.60 (q, J=7.6 Hz, 6H).

Step 4

According to General Method E, TES compound 30 (0.27 g, 0.45 mmol),Me₄NF.4H₂O (0.11 g, 0.68 mmol), AcOH (51 μL, 0.89 mmol) in THF (10 mL)and DMF (2.5 mL) were reacted for 15.5 h to afford the desired OHproduct 31 (0.14 g, 64%).

¹H NMR (CDCl₃, 400 MHz): δ 8.23 (dd, J=8.8, 2.0 Hz, 2H), 7.66 (d, J=8.8Hz, 2H), 5.65 (d, J=7.6 Hz, 1H), 5.49 (d, J=14.0 Hz, 1H), 5.22 (d,J=14.0 Hz, 1H), 4.47-4.38 (m, 1H), 4.32-4.25 (m, 1H), 4.22 (dd, J=10.0,2.8 Hz, 1H), 3.84 (d, J=14.4 Hz, 1H), 3.36 (d, J=14.4 Hz, 1H), 3.36-3.30(m, 1H), 3.27 (dd, J=7.6, 3.2 Hz, 1H), 2.83-2.75 (m, 1H), 2.59 (dd,J=10.0, 6.4 Hz, 1H), 2.49 (dd, J=9.6, 2.8 Hz, 1H), 2.45 (dd, J=14.8, 8.4Hz, 1H), 2.31-2.23 (m, 1H), 1.96 (s, 3H), 1.64-1.53 (m, 2H), 1.37 (d,J=6.0 Hz, 3H), 1.19 (d, J=7.6 Hz, 3H).

Step 5

According to General Method F, OH compound 31 (0.14 g, 0.29 mmol), 10%Pt/C (140 mg) in IPA (5 mL), THF (15 mL), DI water (10 mL) and 0.35 Mphosphate buffer (pH 6, 4 mL) were reacted for 8 h to afford the desiredfinal product 32 (36 mg, 36%).

¹H NMR (D₂O, 400 MHz): δ 4.48-4.39 (m, 1H), 4.24 (t, J=6.4 Hz, 2H),4.15-3.92 (m, 2H), 3.48-3.20 (m, 6H), 2.50-2.35 (m, 1H), 2.04-1.94 (m,1H), 1.97 (s, 3H), 1.28 (d, J=6.4 Hz, 3H), 1.16 (d, J=6.8 Hz, 3H).

Example 6 Synthesis of Compound 37

Step 1

A solution of amine 22 (1.3 g, 7 mmol) in dry THF (70 mL) was cooled to0° C., then ethyl trifluoroacetate (836 μL, 7 mmol) were added into thesolution, aged at 0° C. overnight. After concentration, the residue waspurified by silica gel column chromatography to give the desired amide33 (0.28 g, 14%).

¹H NMR (CDCl₃, 400 MHz): δ 6.49 (d, J=6.0 Hz, 1H), 4.54-4.45 (m, 1H),3.66 (dd, J=11.6, 6.0 Hz, 1H), 3.60-3.20 (m, 3H), 2.28-2.18 (m, 1H),2.05-1.88 (m, 1H), 1.46 (s, 9H).

Step 2

The similar procedure with side chain, 24, synthesis was used forde-protection of Boc group to afford the desired amine 34 in 83% yield.

¹H NMR (CDCl₃, 400 MHz): δ 4.69-4.61 (m, 1H), 3.54-3.45 (m, 1H),3.43-3.33 (m, 1H), 3.30-3.22 (m, 1H), 2.38-2.27 (m, 1H), 2.19-2.09 (m,1H).

Step 3

According to General Method A, CPI (0.43 g, 0.73 mmol), side chain 34(0.13 g, 0.73 mmol), Pd₂(dba)₃CHCl₃ (38 mg, 0.037 mmol) and P(OEt)₃ (42μL, 0.24 mmol) in DMF (18 mL) were reacted for 42 h to afford thedesired TES product 35 (0.3 g, 63%).

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (dd, J=8.8, 2.0 Hz, 2H), 7.65 (d, J=9.2Hz, 2H), 6.69 (br s, 1H), 5.44 (d, J=14.0 Hz, 1H), 5.21 (d, J=14.0 Hz,1H), 4.48-4.42 (m, 1H), 4.27-4.21 (m, 1H), 4.18 (dd, J=10.4, 3.2 Hz,1H), 3.87 (d, J=14.4 Hz, 1H), 3.36 (d, J=14.4 Hz, 1H), 3.27-3.19 (m,2H), 2.91-2.86 (m, 1H), 2.64-2.61 (m, 1H), 2.57-2.53 (m, 1H), 2.47-2.41(m, 1H), 2.36-2.27 (m, 1H), 1.74-1.66 (m, 1H), 1.26 (d, J=6.0 Hz, 3H),1.17 (d, J=7.6 Hz, 3H), 0.93 (t, J=8.0 Hz, 9H), 0.59 (q, J=7.2 Hz, 6H).

Step 4

According to General Method E, TES compound 35 (0.3 g, 0.46 mmol),Me₄NF.4H₂O (0.11 g, 0.68 mmole), AcOH (52 μL, 0.89 mmol) in THF (10 mL)and DMF (2.5 mL) were reacted for 15.5 h to afford the desired OHproduct 36 (0.14 g, 56%).

¹H NMR (CDCl₃, 400 MHz): δ 8.20 (dd, J=8.8, 1.6 Hz, 2H), 7.63 (d, J=8.8Hz, 2H), 6.85 (br s, 1H), 5.45 (d, J=13.6 Hz, 1H), 5.18 (d, J=13.6 Hz,1H), 4.48-4.41 (m, 1H), 4.28-4.21 (m, 2H), 3.87 (d, J=14.4 Hz, 1H), 3.35(d, J=14.0 Hz, 1H), 3.32-3.25 (m, 2H), 2.93-2.87 (m, 1H), 2.65-2.53 (m,3H), 2.44-2.36 (m, 1H), 2.36-2.27 (m, 1H), 1.74-1.66 (m, 1H), 1.32 (d,J=6.4 Hz, 3H), 1.17 (d, J=7.6 Hz, 3H).

Step 5

According to General Method F, OH compound 36 (0.14 g, 0.26 mmol), 10%Pt/C (140 mg) in IPA (5 mL), THF (10 mL), DI water (10 mL) and 0.35 Mphosphate buffer (pH 6, 4 mL) were reacted for 8 h to afford the desiredfinal product 37 (26 mg, 25%).

¹H NMR (D₂O, 400 MHz): δ 4.57-4.48 (m, 1H), 4.25-4.20 (m, 2H), 3.95-3.82(m, 2H), 3.51-3.40 (m, 3H), 3.27-3.18 (m, 2H), 2.50-2.35 (m, 1H),2.10-1.93 (m, 1H), 1.28 (d, J=6.4 Hz, 3H), 1.14 (d, J=6.8 Hz, 3H).

Example 7 Synthesis of Compound 43

Step 1

Mesylate 20 (1.33 g, 5 mmol) and 2 M solution of MeNH₂ in THF (25 mL, 50mmol) were loaded to a sealed tube and aged at 95° C. for 60 h, then thereaction mixture was concentrated and the residue was purified by silicagel column chromatography to give the desired amine 38 (0.85 g, 85%).

¹H NMR (CDCl₃, 400 MHz): δ 3.58-3.28 (m, 3H), 3.26-3.02 (m, 2H), 2.43(s, 3H), 2.08-1.98 (m, 1H), 1.76-1.63 (m, 1H), 1.45 (s, 9H).

Step 2

The similar procedure with side chain, 23, synthesis was used forprotection of nitrogen atom to afford the desired carbamate 39 in 93%yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (d, J=8.8 Hz, 2H), 7.51 (d, J=8.8 Hz,2H), 5.23 (s, 2H), 4.90-4.60 (m, 1H), 3.62-3.44 (m, 2H), 3.38-3.12 (m,2H), 2.88 (s, 3H), 2.10-1.90 (m, 2H), 1.45 (s, 9H).

Step 3

The similar procedure with side chain, 24, synthesis was used forde-protection of Boc group to afford the desired amine 40 in 87% yield.

¹H NMR (DMSO-d₆, 400 MHz): δ 9.12-8.80 (br s, 2H), 8.24 (d, J=8.8 Hz,2H), 7.64 (d, J=8.8 Hz, 2H), 5.24 (s, 2H), 4.82-4.65 (m, 1H), 3.42-3.48(m, 2H), 3.22-3.08 (m, 2H), 2.85 (s, 3H), 2.17-2.06 (m, 1H), 2.03-1.91(m, 1H).

Step 4

According to General Method A, CPI (0.46 g, 0.78 mmol), side chain 40(0.22 g, 0.78 mmol), Pd₂(dba)₃CHCl₃ (41 mg, 0.04 mmol) and P(OEt)₃ (45μL, 0.26 mmol) in DMF (16 mL) were reacted for 24 h to afford thedesired TES product 41 (0.33 g, 56%).

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (dd, J=7.6, 2.0 Hz, 4H), 7.66 (d, J=8.8Hz, 2H), 7.50 (d, J=8.8 Hz, 2H), 5.45 (d, J=14.0 Hz, 1H), 5.22 (d,J=14.0 Hz, 1H), 5.21 (s, 2H), 4.87 (br s, 1H), 4.28-4.22 (m, 1H), 4.18(dd, J=10.0, 3.2 Hz, 1H), 3.77 (d, J=14.4 Hz, 1H), 3.37 (d, J=14.4 Hz,1H), 3.31-3.23 (m, 2H), 2.91 (s, 3H), 2.83-2.78 (m, 1H), 2.64 (br s,1H), 2.46 (t, J=8.4 Hz, 1H), 2.39 (dd, J=16.4, 8.0 Hz, 1H), 2.20-2.09(m, 1H), 1.76 (br s, 1H), 1.26 (d, J=6.0 Hz, 3H), 1.18 (d, J=7.6 Hz,3H), 0.94 (t, J=8.0 Hz, 9H), 0.59 (q, J=8.0 Hz, 6H).

Step 5

According to General Method E, TES compound 41 (0.33 g, 0.44 mmol),Me₄NF.4H₂O (0.11 g, 0.68 mmol), AcOH (50 μL, 0.87 mmol) in THF (10 mL)and DMF (2.5 mL) were reacted for 16 h to afford the desired OH product42 (0.18 g, 64%).

¹H NMR (CDCl₃, 400 MHz): δ 8.20 (d, J=8.4 Hz, 4H), 7.64 (d, J=8.4 Hz,2H), 7.49 (d, J=8.0 Hz, 2H), 5.47 (d, J=13.6 Hz, 1H), 5.29 (s, 2H), 5.20(d, J=13.6 Hz, 1H), 4.81 (br s, 1H), 4.27-4.20 (m, 2H), 3.76 (d, J=14.4Hz, 1H), 3.36 (d, J=14.4 Hz, 1H), 3.32-3.27 (m, 2H), 2.91 (s, 3H),2.83-2.79 (m, 1H), 2.45 (t, J=8.8 Hz, 1H), 2.38 (dd, J=16.4, 8.0 Hz,1H), 2.13 (br s, 2H), 1.76 (br s, 1H), 1.34 (d, J=6.0 Hz, 3H), 1.19 (d,J=7.2 Hz, 3H).

Step 6

According to General Method F, OH compound 42 (0.21 g, 0.33 mmol), 10%Pt/C (300 mg) in IPA (5 mL), THF (10 mL), DI water (10 mL) and 0.35 Mphosphate buffer (pH 6, 4 mL) were reacted for 8 h to afford the desiredfinal product 43 (29 mg, 27%).

¹H NMR (D₂O, 400 MHz): δ 4.25-4.18 (m, 2H), 3.74 (d, J=13.6 Hz, 1H),3.68-3.61 (m, 1H), 3.48 (d, J=13.6 Hz, 1H), 3.43 (dd, J=6.0, 2.8 Hz,2H), 3.25-3.18 (m, 1H), 3.12-3.06 (m, 1H), 2.81 (br s, 1H), 2.75-2.69(m, 1H), 2.67-2.61 (m, 1H), 2.58 (s, 3H), 2.34-2.24 (m, 1H), 1.90-1.82(m, 1H), 1.28 (d, J=6.0 Hz, 3H), 1.11 (d, J=7.6 Hz, 3H).

Example 8 Synthesis of Compound 46

Step 1

The similar procedure with side chain, 38, synthesis was used to affordthe desired dimethyl amine 43 in 64% yield.

¹H NMR (CDCl₃, 400 MHz): δ 3.70-3.61 (m, 0.5H), 3.60-3.43 (m, 2H),3.32-3.21 (m, 1H), 2.72-2.56 (m, 1.5H), 2.26 (s, 6H), 2.09-2.01 (m, 1H),1.80-1.67 (m, 1H), 1.46 (s, 9H).

Step 2

The similar procedure with side chain, 24, synthesis was used forde-protection of Boc group to afford the desired amine 44 as a TFA saltin quantitative yield.

¹H NMR (CDCl₃, 400 MHz): δ 4.10 (m, 1H), 3.82 (dd, J=12.4, 8.4 Hz, 1H),3.62 (dd, J=12.4, 7.6 Hz, 1H), 3.52 (ddd, J=11.6, 8.4, 4.0 Hz, 1H), 3.40(ddd, J=11.6, 10.0, 7.6 Hz, 1H), 2.88 (s, 6H), 2.54-2.44 (m, 1H),2.35-2.23 (m, 1H).

Step 3

According to General Method B, CPI (0.77 g, 1.3 mmol), side chain 44(0.28 g, 1.3 mmol), Pd₂(dba)₃CHCl₃ (68 mg, 0.066 mmol), P(OEt)₃ (75 μL,0.43 mmol) and 2,6-lutidine (0.3 mL, 2.58 mmol) in DMF (23 mL) werereacted for 74.5 h to afford the OH product 45 (0.17 g, 27%).

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (d, J=8.8 Hz, 2H), 7.64 (d, J=8.8 Hz,2H), 5.46 (d, J=13.6 Hz, 1H), 5.19 (d, J=14.0 Hz, 1H), 4.29-4.21 (m,2H), 3.89 (d, J=13.6 Hz, 1H), 3.68 (br s, 1H), 3.40 (d, J=14.4 Hz, 1H),3.32 (t, J=8.4 Hz, 1H), 3.26 (dd, J=6.4, 2.8 Hz, 1H), 3.12 (d, J=8.8 Hz,1H), 3.01-2.97 (m, 1H), 2.75 (s, 6H), 2.52 (br s, 2H), 2.25-2.18 (m,1H), 2.12-2.03 (m, 1H), 1.33 (d, J=6.0 Hz, 3H), 1.15 (d, J=7.2 Hz, 3H).

Step 4

According to General Method F, OH compound 45 (0.12 g, 0.25 mmol), 10%Pt/C (120 mg) in IPA (4 mL), THF (8 mL), DI water (8 mL) and 0.35 Mphosphate buffer (pH=6, 3 mL) were reacted for 7 h to afford the desiredfinal product 46 (20 mg, 23%).

¹H NMR (D₂O, 400 MHz): δ 4.28-4.14 (m, 2H), 3.86-3.75 (m, 1H), 3.62-3.43(m, 3H), 3.17-3.11 (m, 2H), 3.02-2.75 (m, 3H), 2.55 (s, 6H), 2.33-2.20(m, 1H), 2.01-1.86 (m, 1H), 1.26 (d, J=6.0 Hz, 3H), 1.11 (d, J=7.6 Hz,3H).

Example 9 Synthesis of Compound 49

Step 1

According to General Method C, CPI (3.48 g, 5.89 mmol),3-azetidinecarboxylic amide (0.59 g, 5.89 mmol), Pd₂(dba)₃CHCl₃ (0.3 g,0.29 mmol), P(OEt)₃ (0.34 mL, 1.95 mmol) and TsOH (0.56 g, 2.94 mmol) inDMF (100 mL) were reacted for 91 h to afford the desired TES product 47(1.03 g, 31%).

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (d, J=8.8 Hz, 2H), 7.67 (d, J=8.8 Hz,2H), 6.04 (br s, 1H), 5.46 (d, J=13.6 Hz, 1H), 5.39 (br s, 1H), 5.23 (d,J=13.6 Hz, 1H), 4.27-4.21 (m, 1H), 4.18 (dd, J=10.4, 2.8 Hz, 1H), 3.98(d, J=14.4 Hz, 1H), 3.50-3.36 (m, 4H), 3.27-3.19 (m, 3H), 3.13-3.06 (m,1H), 1.25 (d, J=6.4 Hz, 3H), 1.16 (d, J=7.2 Hz, 3H), 0.94 (t, J=8.0 Hz,9H), 0.59 (q, J=7.6 Hz, 6H).

Step 2

According to General Method E, TES compound 47 (0.8 g, 1.4 mmol),Me₄NF.4H₂O (0.41 g, 2.5 mmol), AcOH (0.2 mL, 3.5 mmol) in THF (30 mL)and DMF (10 mL) were reacted for 15.5 h to afford the desired OH product48 (0.35 g, 55%).

¹H NMR (CDCl₃, 400 MHz): δ 8.23 (d, J=8.8 Hz, 2H), 7.66 (d, J=8.8 Hz,2H), 6.08 (br s, 1H), 5.55 (br s, 1H), 5.49 (d, J=13.6 Hz, 1H), 5.22 (d,J=13.6 Hz, 1H), 4.29-4.24 (m, 1H), 4.20 (dd, J=10.0, 3.2 Hz, 1H), 3.97(d, J=14.4 Hz, 1H), 3.48-3.34 (m, 4H), 3.32-3.21 (m, 3H), 3.13-3.06 (m,1H), 1.34 (d, J=6.4 Hz, 3H), 1.16 (d, J=7.2 Hz, 3H).

Step 3

According to General Method F, OH compound 48 (0.35 g, 1.18 mmol), 5%Pt/C (300 mg) in IPA (10 mL), THF (20 mL), DI water (20 mL) and 0.35 Mphosphate buffer (pH 6, 8 mL) were reacted for 23 h to afford thedesired final product 49 (107 mg, 43%).

¹H NMR (D₂O, 400 MHz): δ 4.19-4.08 (m, 6H), 3.96 (br s, 1H), 3.95-3.85(m, 1H), 3.58 (t, J=8.0 Hz, 1H), 3.40-3.39 (m, 1H), 3.16-3.08 (m, 1H),1.21 (d, J=6.4 Hz, 3H), 1.09 (d, J=7.2 Hz, 3H).

Example 10 Synthesis of Compound 58

Step 1

3-Hydroxyazetidine hydrochloride (10.96 g, 0.1 mol) was dissolved in H₂O(20 ml) and CH₂Cl₂ (200 mL) and cooled with ice-bath to 0° C. To theabove solution was added NaHCO₃ (8.4 g, 0.1 mmol) slowly as a solid insmall portions, aged at 0° C. for 10 min. After the addition, then Et₃N(20.9 mL, 0.15 mole) was added, followed by dropwise addition ofsolution of (Boc)₂O (24 g, 0.11 mol) in CH₂Cl₂ (30 mL), kept stirring at0° C. for 1 h. The reaction mixture was treated with H₂O (200 mL),stirred for 10 min, and separated. The aqueous phase was extracted withCH₂Cl₂ (100 mL) twice, and the combined organic layers was washed withbrine (200 mL), dried over anhydrous Na₂SO₄ and filtered. The filtratewas concentrated and dried over high vacuum to give the desiredcarbamate 50 as an oily product (crude 19.3 g, used for next stepwithout further purification).

¹H NMR (CDCl₃, 400 MHz): δ 4.60-4.50 (m, 1H), 4.10 (ddd, J=9.6, 6.8, 0.8Hz, 2H), 3.78 (ddd, J=9.6, 4.4, 0.8 Hz, 2H), 1.42 (s, 9H).

Step 2

The carbamate 50 (4.36 g, 25.2 mmol) was dissolved in dry CH₂Cl₂ (200mL) and cooled with ice-bath to 0° C. To the above solution was addedEt₃N (7 mL, 50.34 mmol), followed by dropwise addition of MsCl (2.54 mL,32.72 mmol). After 2 h at 0° C., the reaction mixture was treated withH₂O (100 mL), stirred for 10 min, and separated. The aqueous phase wasextracted with CH₂Cl₂ (50 mL) twice, and the combined organic layers waswashed with brine (100 mL), dried over anhydrous Na₂SO₄ and filtered.The filtrate was concentrated and dried over high vacuum to give themesylate 51 as an oily product (crude 6.38 g, used for next step withoutfurther purification)

¹H NMR (CDCl₃, 400 MHz): δ 5.22-5.16 (m, 1H), 4.26 (ddd, J=10.4, 6.8,1.2 Hz, 2H), 4.08 (ddd, J=10.4, 4.4, 1.2 Hz, 2H), 3.06 (s, 3H), 1.42 (s,9H).

Step 3

The similar procedure with side chain, 21, synthesis was used forazidation to afford the desired azide 52 in 92% yield.

¹H NMR (CDCl₃, 400 MHz): δ 4.24-4.16 (m, 3H), 3.91-3.86 (m, 2H), 1.43(s, 9H).

Step 4

The similar procedure with side chain, 22, synthesis was used for thereduction of azide to afford the desired amine 53 in quantitative yield.

¹H NMR (CDCl₃, 400 MHz): δ 4.13 (dd, J=8.4, 8.0 Hz, 2H), 3.80-3.70 (m,1H), 3.56 (dd, J=9.2, 5.2 Hz, 2H), 1.42 (s, 9H).

Step 5

To a 200 mL oven-dried round-bottomed flask charged with amine 53 (0.69g, 4.0 mmol) was added dry 1,4-dioxane (40 mL) to give a colorlesssolution under N₂. Then sulfamide (0.77 g, 8.0 mmol) was added. Thismixture was put into a pre-heated oil-bath (85° C.). After heating for52 h, oil-bath was removed. The crude mixture was concentrated in vacuo.The residue was treated with DI water and DCM. The aqueous layer wasseparated and extracted with DCM (×5). The combined organic layer wasdried (Na₂SO₄) and concentrated in vacuo. The crude material waspurified by SiO₂ column chromatography eluting from 1% to 3% of MeOH inDCM to provide the desired product 54 (0.34 g, 34%).

¹H NMR (CDCl₃, 400 MHz): δ 5.73 (br s, 1H), 5.20 (br s, 1H), 5.16 (br s,1H), 4.25-4.22 (m, 3H), 3.89 (d, J=4.8 Hz, 2H), 1.81 (br s, 1H), 1.43(s, 9H).

Step 6

The similar procedure with side chain, 24, synthesis was used forde-protection of Boc group to afford the desired amine 55 as a TFA saltin quantitative yield.

¹H NMR (DMSO-d6, 400 MHz): δ 8.74 (br s, 1H), 8.66 (br s, 1H), 7.52 (d,J=8.0 Hz, 1H), 6.85 (s, 2H), 4.25-4.16 (m, 1H), 4.08-4.07 (m, 2H),3.90-3.87 (m, 2H).

Step 7

According to General Method B, CPI (0.80 g, 1.35 mmol), side chain 55(0.38 g, 1.35 mmol), Pd₂(dba)₃CHCl₃ (70 mg, 0.068 mmol), P(OEt)₃ (78 μL,0.45 mmol) and 2,6-lutidine (0.31 mL, 2.67 mmol) in DMF (23 mL) werereacted for 73 h to afford the desired TES product 56 (0.24 g, 29%).

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (d, J=8.8 Hz, 2H), 7.67 (d, J=8.8 Hz,2H), 5.66 (br s, 1H), 5.44 (d, J=13.6 Hz, 1H), 5.29 (d, J=14.0 Hz, 1H),4.73 (d, J=13.3 Hz, 1H), 4.40-4.01 (m, 10H), 3.79-3.62 (m, 3H),3.35-3.28 (m, 2H), 1.22 (d, J=6.0 Hz, 3H), 1.14 (d, J=6.8 Hz, 3H), 0.92(t, J=8.0 Hz, 9H), 0.58 (t, J=7.2 Hz, 6H).

Step 8

According to General Method E, TES compound 56 (0.24 g, 0.38 mmol),Me₄NF.4H₂O (0.11 g, 0.68 mmol), AcOH (56 μL, 0.98 mmol) in THF (10 mL)and DMF (3 mL) were reacted for 18.5 h to afford the desired OH product57 (0.17 g, 87%).

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (d, J=8.4 Hz, 2H), 7.65 (d, J=8.4 Hz,2H), 5.46 (d, J=13.6 Hz, 1H), 5.22 (d, J=13.6 Hz, 1H), 4.23-4.17 (m,2H), 4.04-4.00 (m, 1H), 3.95 (d, J=14.4 Hz, 1H), 3.67-3.60 (m, 2H), 3.43(d, J=14.8 Hz, 1H), 3.27-3.21 (m, 3H), 3.10 (t, J=6.8 Hz, 1H), 3.02 (t,J=6.8 Hz, 1H), 1.31 (d, J=6.0 Hz, 3H), 1.13 (d, J=7.2 Hz, 3H).

Step 9

According to General Method F, OH compound 57 (0.17 g, 0.33 mmol), 5%Pt/C (270 mg) in IPA (4.5 mL), THF (9 mL), DI water (9 mL) and 0.35 Mphosphate buffer (pH 6, 4 mL) were reacted for 8 h to afford the desiredfinal product 58 (19 mg, 15%).

¹H NMR (D₂O, 400 MHz): δ 4.28 (br s, 2H), 4.21-4.13 (m, 4H), 3.87 (br s,2H), 3.72-3.61 (m, 1H), 3.41-3.39 (m, 1H), 3.17-3.06 (m, 1H), 1.22 (d,J=6.0 Hz, 3H), 1.08 (d, J=7.2 Hz, 3H).

Example 11 Synthesis of Compound 64

Step 1

N,N′-bis(p-nitrobenzyloxycarbonyl)-S-methylisothiourea 59 wassynthesized as described in U.S. Patent Publication No. 2005-020519 andWO 2005/123069 A02. To a solution of compound 22 (18.7 g, 100 mmol) inTHF (1 L) was added methylisothiourea 59 (39.05 g, 80 mmol), agedovernight at rt and then concentrated down to about 200 mL volume. Theresidue was triturated with MeOH (200 mL) and concentrated to about 200mL volume again (repeated the trituration and concentration twice). Theprecipitated solid was filtered off, washed with MeOH (50 mL) twice anddried overnight at high vacuum to give the desired guanidine 60 (40 g,80%) as a white solid.

¹H NMR (CDCl₃, 400 MHz): δ 11.76 (s, 1H), 8.42 (s, 1H), 8.23 (dd,J=15.2, 8.8 Hz, 4H), 7.54 (dd, J=8.8, 7.2 Hz, 4H), 5.27 (s, 2H), 5.22(s, 2H), 4.72-4.60 (m, 1H), 3.72-3.60 (m, 1H), 3.52-3.38 (m, 2H),3.26-3.16 (m, 1H), 2.26-2.16 (m, 1H), 1.95-1.84 (m, 1H), 1.45 (s, 9H).

Step 2

To a solution of TFA (52.1 mL, 677 mmol) in CH₂Cl₂ (250 mL) at 0° C. wasadded the guanidine 60 (28.25 g, 45.13 mmol) as a solid, the reactionmixture was aged at 0° C. overnight, then it was concentrated and theresidue was purified by silica gel column chromatography. The fractionsare collected and concentrated and then triturated with EtOAc (50 mL)twice. The precipitated solid was collected by filtration, washed thecake with EtOAc (30 mL) twice and dried under high vacuum to afford thedesired amine TFA salt 61 (20 g, 70%) as a white solid.

¹H NMR (CDCl₃, 400 MHz): δ 9.48-9.28 (br s, 1H), 9.20-9.00 (br s, 1H),8.22 (dd, J=16.8, 8.8 Hz, 4H), 7.53 (dd, J=8.8, 6.4 Hz, 4H), 5.60-5.50(br s, 2H), 5.30 (s, 2H), 5.21 (d, J=1.6 Hz, 2H), 4.61-4.53 (m, 1H),3.80-3.68 (m, 1H), 3.66-3.54 (m, 2H), 3.44-3.30 (m, 1H), 2.62-2.48 (m,1H), 2.32-2.18 (m, 1H).

Step 3 & 4

In a degassed DMF (20 mL), Pd₂(dba)₃-CHCl₃(76 mg, 0.073 mmol) andtriethyl phosphite (78 μL, 0.454 mmol) were added and mixed a catalystuntil forming a deep yellow solution at rt. To the catalytic solutionwas added CPI (400 mg, 0.67 mmol), amine•TFA salt 61 (350 mg, 0.58mmole) and 2,6-lutidine (200 μL, 2.96 mmol) and the resulting mixturewas stirred 2 days at rt. After concentration under vacuum, the mixturewas purified with 65% Ethyl acetate in Hexane to afford a 1:1 mixture of62 & 63 (670 mg). The mixture was dissolved in THF and DMF (15 mL/5 mL)and followed by addition of acetic acid (200 μL, 3.5 mmol) andMe₄NF.4H₂O (200 mg, 1.2 mmol) at 4° C. After stirring overnight, themixture was quenched with sat. NaHCO₃ and extracted with DCM twice. Theextract was dried over anhydrous MgSO₄ and concentrated under a reducedpressure. The concentrate was purified by a silica column chromatographyto afford the pure alcohol 63 (420 mg, 86% over two-steps).

¹H NMR (CDCl₃, 400 MHz): δ 11.73 (s, 1H), 8.54 (d, J=8.0 Hz, 1H), 8.2(m, 6H), 7.63 (d, J=9.2 Hz, 2H), 7.52 (d, J=8.8 Hz, 2H), 7.51 (d, J=8.8Hz, 2H), 5.46 (d, J=14.0 Hz, 1H), 5.23 (s, 2H), 5.19 (s, 2H), 5.18 (d,J=14.4 Hz, 1H) 4.60 (m, 1H), 4.21 (m, 2H), 3.82 (d, J=14.4 Hz, 2H), 3.37(d, J=14.4 Hz, 1H), 3.36 (m, 1H), 3.26 (dd, J=5.8, 3.2 Hz, 1H), 2.57(dd, J=9.6, 2.4 Hz, 2H), 2.50 (dd, J=9.6, 5.6 Hz, 1H), 2.40 (q, J=5.8Hz, 1H), 2.26 (m, 1H), 1.90 (m, 1H), 1.69 (m, 1H), 1.30 (d, J=7.2 Hz,3H), 1.18 (d, J=7.2 Hz, 3H)

Step 5

According to General Method F, OH compound 63 (0.37 g, 0.438 mmol), 5%Pt/C (270 mg) in IPA (5 mL), THF (10 mL), and 0.35 M phosphate buffer(pH 6, 10 mL) were reacted for 7 h to afford the desired final product64 (25 mg, 16.3%).

¹H NMR (D₂O, 400 MHz): δ 4.08 (m, 1H), 4.02 (dd, J=9.6, 2.8 Hz, 1H),3.54 (d, J=13.2 Hz, 1H), 3.26 (dd, J=5.4, 2.8 Hz, 1H), 3.22 (d, J=13.6Hz, 1H), 3.68 (dd, J=11.2, 7.2 Hz, 1H), 2.60 (m, 1H), 2.46 (m, 2H), 2.17(m, 1H), 2.63 (m, 1H), 1.12 (d, J=6.0 Hz, 3H), 0.95 (d, J=7.6 Hz, 3H).

Example 12 Synthesis of Compound 67

Step 1

To a solution of amine 62 (1.39 g, 1.45 mmol) in THF (7 mL) was added350 μL of MeI (5.62 mmol) at 0° C. and warmed up to rt. After 3 days itwas concentrated and washed with a saturated brine to exchange thecounter anion. The extract was dried over anhydrous MgSO₄ andconcentrated under a reduced pressure. The concentrate was purified by asilica column chromatography with 10% MeOH in DCM to afford thequarternary amine salt 65 (1 g, 49% yield).

¹H NMR (Acetone-d6/CDCl₃, 400 MHz): δ 11.53 (s, 1H), 8.66 (d, J=6.8 Hz,0.6H), 8.49 (d, J=5.6 Hz, 0.4H), 8.10 (m, 6H), 7.62-7.45 (m, 6H),5.40-4.87 (m, 6H) 4.36-1.91 (m, 13H), 3.38 (s, 0.4H), 3.36 (s, 0.6H),1.28-1.05 (m, 6H), 0.84 (m, 9H), 0.49 (q, J=8.0 Hz, 6H).

Step 2

According to General Method E, TES compound 65 (1.0 g, 1.0 mmol),Me₄NF.4H₂O (0.25 g, 1.5 mmol), AcOH (200 μL, 3.5 mmol) in THF (15 mL)and DMF (5 mL) were reacted overnight to afford the desired OH product66 (0.40 g, 45%).

¹H NMR (CD₃OD/CDCl₃, 400 MHz): δ 8.22-8.15 (m, 6H), 7.63 (dd, J=5.6, 4.4Hz, 2H), 7.53-7.47 (m, 4H), 5.43-5.05 (m, 6H), 4.95-4.80 (2 m, 1H), 4.43(m, 2H), 4.20-3.95 (m, 3H), 3.75-3.45 (m, 2H), 3.34 (s, 1.8H), 3.26 (m,1H), 3.12 (s, 1.2H), 2.88 (m, 2H), 2.68 (m, 1H), 2.45 (m, 1H), 1.29-1.19(m, 6H).

Step 3

According to General Method G, OH compound 66 (0.18 g, 0.201 mmol), Zincdust (2.77 g) in THF (10 mL) and 0.35 M phosphate buffer (pH 6, 20 mL)were reacted for 7 h to afford the desired final product 67 (9 mg,12.3%).

¹H NMR (D₂O, 400 MHz): δ 4.90 (m, 1H), 4.40 (br s, 1H), 4.15 (m, 2H),3.78 (m, 1H), 3.60 (m, 2H), 3.45 (m, 1H), 3.35 (br s, 1H), 3.25-3.05 (m,2H), 3.02 (s, 1.2H), 3.85 (s, 1.8H), 2.55 (m, 1H), 2.06 (m, 1H), 1.10(m, 3H), 0.92 (m, 3H).

Example 13 Synthesis of Compound 72

Step 1

The similar procedure with side chain, 54, synthesis was used for thesubstitution reaction to afford the desired sulfonamide 68 in 24% yield.

¹H NMR (CDCl₃, 400 MHz): δ 5.29-4.94 (m, 3H), 4.05-3.95 (m, 1H),3.70-3.55 (m, 1H), 3.53-3.25 (m, 3H), 2.22-2.11 (m, 1H), 2.05-1.90 (m,1H), 1.45 (s, 9H).

Step 2

The similar procedure with side chain, 24, synthesis was used for thedeprotection of Boc group to afford a desired amine 69 as a TFA salt inquantitative yield.

¹H NMR (CD₃OD, 400 MHz): δ 4.14-4.09 (m, 1H), 3.46-3.32 (m, 4H),2.34-2.23 (m, 1H), 2.14-2.05 (m, 1H).

Step 3 & 4

The similar procedures (coupling & deprotection) with the synthesis ofcarbapenem 63 were used to afford the desired carbapenem 71 in 42% yieldover two-steps.

¹H NMR (Acetone-D₆, 400 MHz,): δ 8.25 (d, J=8.8 Hz, 2H), 7.82 (d, J=9.2Hz, 2H), 5.99 (br s, 1H), 5.55 (d, J=14.0 Hz, 1H), 5.34 (d, J=14.4 Hz,1H), 4.28 (dd, J=8.4, 2.8 Hz, 1H), 4.15 (p, J=6.0 Hz, 1H), 4.05 (m, 1H),3.66-3.48 (m, 2H), 3.34 (dd, J=6.4, 2.4 Hz, 1H), 3.27-2.63 (m, 6H), 2.32(br s, 1H), 1.89 (br s, 1H), 1.26 (d, J=6.8 Hz, 3H), 1.21 (d, J=7.6 Hz,3H).

Step 5

According to General Method F, OH compound 71 (0.23 g, 0.438 mmol), 5%Pt/C (270 mg) in IPA (6 mL), THF (12 mL), and 0.35 M phosphate buffer(pH 6, 12 mL) were reacted for 7 h to afford the desired final product72 (30 mg, 18%).

¹H NMR (D₂O in buffer at pH 7, 400 MHz): δ 4.10-3.91 (m, 4H), 3.59-3.40(m, 6H), 3.07 (m, 1H), 2.35 (m, 1H), 1.97 (m, 1H), 1.11 (d, J=6.8 Hz,3H), 1.01 (d, J=7.2 Hz, 3H).

Example 14 Synthesis of Compound 78

Step 1

A solution of compound 22 (0.8 g, 4.3 mmol) in dry CH₂Cl₂ (40 mL) wascooled to 0° C., then DIEA (1.5 mL, 8.6 mmol) and ClCO₂Ph (630 μL, 5mmol) were added into the solution, slowly warm up to rt. Afterovernight, the mixture was treated with H₂O (20 mL), separated andextracted with CH₂Cl₂ (20 mL) twice. The combined organic layers werewashed with brine (30 mL), concentrated and purified by silica gelcolumn chromatography to give the desired carbamate 73 (1.2 g, 91%).

¹H NMR (CDCl₃, 400 MHz): δ 7.36 (dd, J=8.0, 7.6 Hz, 2H), 7.21 (dd,J=7.6, 7.2 Hz, 1H), 7.12 (d, J=8.0 Hz, 2H), 5.13 (d, J=6.8 Hz, 1H),4.36-4.27 (m, 1H), 3.65 (dd, J=11.6, 6.0 Hz, 1H), 3.55-3.39 (m, 2H),3.37-3.22 (m, 1H), 2.26-2.14 (m, 1H), 2.03-1.85 (m, 1H), 1.47 (s, 9H).

Step 2

The carbamate 73 (918 mg, 3 mmol) and 30 mL of NH₃ (7 M solution inMeOH, 210 mmol) were loaded to a sealed tube and aged at 90° C. for 60h. After cooling down to rt, the reaction mixture was concentrated andthe residue was triturated with EtOAc (10 mL×5). The precipitated solidwas collected by filtration, washed the cake with CH₂Cl₂ (5 mL) twiceand dried under high vacuum to give the desired urea 74 (0.547 g, 80%).

¹H NMR (DMSO-d₆, 400 MHz): δ 6.21 (s, 1H), 5.41 (s, 2H), 4.02-3.92 (m,1H), 3.41-3.17 (m, 3H), 2.96 (dd, J=10.8, 4.4 Hz, 1H), 2.00-1.88 (m,1H), 1.71-1.58 (m, 1H), 1.37 (s, 9H).

Step 3

The similar procedure with side chain, 24, synthesis was used fordeprotection of Boc group to afford the desired amine 75 as a TFA saltin quantitative yield.

¹H NMR (DMSO-d₆, 400 MHz): δ 8.79-8.73 (br, 2H), 6.38 (d, J=6.0 Hz, 1H),5.80-5.40 (br s, 2H), 4.13-4.04 (m, 1H), 3.30-3.19 (m, 2H), 3.19-3.09(m, 1H), 2.99-2.90 (m, 1H), 2.11-2.02 (m, 1H), 1.77-1.68 (m, 1H).

Step 4

According to General Method A, CPI (0.59 g, 1.0 mmol), side chain 75(0.243 g, 1.0 mmol), Pd₂(dba)₃CHCl₃ (76 mg, 0.073 mmol) and P(OEt)₃ (78μL, 0.454 mmol) in DMF (20 mL) were reacted overnight to afford thedesired TES product 76 (0.30 g, 50%).

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (d, J=8.8 Hz, 2H), 7.65 (d, J=8.8 Hz,2H), 5.43 (d, J=13.6 Hz, 1H), 5.21 (d, J=14.0 Hz, 1H), 4.78 (m, 1H),4.28-4.11 (m, 3H), 3.86 (d, J=14.4 Hz, 1H), 3.34 (d, J=14.4 Hz, 1H),3.31-3.22 (m, 2H), 2.82 (m, 1H), 2.58 (m, 2H), 2.41 (q, J=8.4 Hz, 1H),2.24 (m, 1H), 1.66 (m, 1H), 1.24 (d, J=6.4 Hz, 3H), 1.15 (d, J=7.2 Hz,3H), 0.93 (t, J=7.6 Hz, 9H), 0.59 (d, J=7.6 Hz, 6H)

Step 5

The general method E was used for deprotection of TES group to affordthe desired OH compound 77 in 85% yield.

¹H NMR (CD₃OD/CDCl₃, 400 MHz): δ 8.14 (d, J=6.8 Hz, 2H), 7.57 (d, J=6.8Hz, 2H), 5.38 (d, J=14.0 Hz, 1H), 5.16 (d, J=13.6 Hz, 1H), 4.09 (m, 2H),3.86 (d, J=13.2 Hz, 1H), 3.31-3.21 (m, 3H), 3.15 (dd, J=2.8, 7.2 Hz,1H), 2.53 (br s, 2H), 2.38 (m, 1H), 2.18 (m, 1H), 1.24 (d, J=6.8 Hz,3H), 1.08 (d, J=7.2 Hz, 3H).

Step 6

The general method F was used for deprotection ofp-nitrobenzyloxycarbonyl group to afford the desired final product 78 in33% yield.

¹H NMR (D₂O, 400 MHz): δ 4.16 (br s, 1H), 4.07 (m, 2H), 3.83 (br s, 2H),3.31 (dd, J=6.0, 2.8 Hz, 1H), 3.35-3.02 (m, 5H), 2.26 (br s, 1H), 1.81(m, 1H), 1.12 (d, J=6.4 Hz, 3H), 1.01 (d, J=7.2 Hz, 3H).

Example 15 Synthesis of Compound 83

Step 1

A solution of compound 22 (1.12 g, 6 mmol) in dry CH₃CN (20 mL) wascooled to 0° C., then Et₃N (1.4 mL, 10 mmol) was added to the reactionmixture followed by bromoacetamide (0.69 g, 5 mmol), and slowly warmedup to rt overnight. After removing the solvent, the residue was treatedwith H₂O (20 mL) and CH₂Cl₂ (50 mL) and separated. The aqueous layer wasextracted by CH₂Cl₂ (20 mL) twice, and the combined organic layers waswashed with brine (40 mL) and concentrated. The concentrate was purifiedby a silica gel column chromatography to give the desiredmono-alkylation product 79 (0.55 g, 45%).

¹H NMR (DMSO-d₆, 400 MHz): δ 7.22 (s, 1H), 7.05 (s, 1H), 3.32-3.22 (m,2H), 3.22-3.08 (m, 2H), 2.98-3.04 (m, 2H), 2.95 (dd, J=10.4, 4.4 Hz,1H), 2.30-2.20 (m, 1H), 1.91-1.80 (m, 1H), 1.67-1.54 (m, 1H), 1.36 (s,9H).

Step 2

The similar procedure with side chain, 24, synthesis was used forde-protection of Boc group to afford the desired amine 80 as a TFA saltin quantitative yield.

¹H NMR (CDCl₃, 400 MHz): δ 3.34-3.28 (m, 1H), 3.26-3.18 (m, 2H),3.10-3.00 (m, 3H), 2.95 (d, J=4.8 Hz, 2H), 2.18-1.86 (m, 1H), 1.74-1.65(m, 1H).

Step 3

According to General Method B, CPI (0.59 g, 1.0 mmol), side chain 80(0.30 g, 2.1 mmol), Pd₂(dba)₃CHCl₃ (76 mg, 0.073 mmol), P(OEt)₃ (78 μL,0.454 mmol) and 2,6-lutidine (0.232 mL, 2.0 mmol) in DMF (20 mL) werereacted for 4 h to afford the desired coupling product 81 (0.30 g, 49%).

¹H NMR (CDCl₃, 400 MHz): δ 8.20 (d, J=8.8 Hz, 2H), 7.65 (d, J=8.8 Hz,2H), 7.20 (br s, 1H), 6.00 (br s, 1H), 5.43 (d, J=14.0 Hz, 1H), 5.21 (d,J=14.4 Hz, 1H), 4.28-4.06 (m, 2H), 3.92 (m, 1H), 3.39-3.23 (m, 5H), 2.79(br s, 1H), 2.58 (br s, 2H), 2.11 (m, 1H), 1.62 (br s, 1H), 1.23 (d,J=6.0 Hz, 3H), 1.15 (d, J=7.2 Hz, 3H), 0.92 (t, J=8.0 Hz, 9H), 0.56 (q,J=8.0 Hz, 6H).

Step 4

The general method E was used for deprotection of TES group to affordthe desired OH compound 82 in 49% yield.

¹H NMR (Acetone-d6/CDCl₃, 400 MHz): δ 8.08 (br s, 2H), 7.55 (d, J=6.4Hz, 2H), 7.14 (br s, 1H), 6.19 (br s, 1H), 5.36 (d, J=14.0 Hz, 1H), 5.09(d, J=13.6 Hz, 1H), 4.10 (br s, 1H), 3.73 (d, J=13.2 Hz, 1H), 3.52 (brs, 1H), 3.27-3.05 (m, 6H), 2.69 (br s, 1H), 2.45 (br s, 1H), 2.32 (br s,2H), 1.99 (br s, 1H), 1.47 (br s. 1H), 1.20 (br s, 3H), 1.03 (br s, 3H)

Step 5

The general method F was used for deprotection ofp-nitrobenzyloxycarbonyl group to afford the desired final product 83 in16% yield.

¹H NMR (D₂O, 400 MHz): δ 4.04-4.00 (m, 2H), 3.88-3.71 (m, 3H), 3.36 (brs, 1H), 3.27 (dd, J=2.8, 6.0 Hz, 1H), 3.13 (m, 1H), 3.10 (d, J=11.2 Hz,2H), 3.05-2.90 (m, 3H), 2.55 (br s, 1H), 1.68 (br s, 1H), 1.06 (d, J=6.4Hz, 3H), 0.95 (d, J=7.2 Hz, 3H).

Example 16 Synthesis of Compound 84

To a iso-propyl formimidate HCl salt (618 mg, 5 mmole) in iso-propanol(12 mL) was added 870 μL of DIEA (5 mmole) at −15° C. After 10 minute,it was transferred into a buffer solution (pH 7, 0.25 M, 25 mL) of amine27 (100 mg, 0.32 mmole) at ice-bath and stirred over 3 hrs. The mixturewas diluted cold DI water (25 mL) and washed with cold ethyl acetatetwice. The aqueous layer was lyophilized and then purified on SP-207resin with a solvent gradient system (from 100% water to 45% i-PrOH inwater). The column fractions containing product were then concentratedunder vacuum and lyophilized to afford the desired amidine carbapenem 84(19 mg, 17.5%).

¹H NMR (D₂O, 400 MHz): δ 7.66 (s, 0.3H), 7.55 (s, 0.7H), 4.15 (m, 1H),4.04-3.98 (m, 2H), 3.63-3.42 (m, 3H), 3.24 (m, 1H), 3.03-2.66 (m, 4H),2.27 (m, 1H), 1.77 (m, 1H), 1.12 (d, J=6.4 Hz, 0.9H), 1.07 (d, J=6.4 Hz,2.1H), 0.95 (d, J=7.2 Hz, 0.9H), 0.92 (d, J=7.2 Hz, 2.1H).

Example 17 Synthesis of Compound 89

Step 1

The similar procedure with side chain, 60, synthesis was used forguanidation to afford the desired guanidine 85 in 70% yield.

¹H NMR (CDCl₃, 400 MHz): δ 11.73 (s, 1H), 8.66 (d, J=6.4 Hz, 1H), 8.24(dd, J=8.8, 17.2 Hz, 4H), 7.54 (dd, J=8.8, 6.4 Hz, 4H), 5.29 (s, 2H),5.21 (s, 2H), 4.78-4.68 (m, 1H), 4.28 (dd, J=9.6, 7.6 Hz, 2H), 3.79 (dd,J=9.6, 5.2 Hz, 2H), 1.43 (s, 9H).

Step 2

The similar procedure with side chain, 61, synthesis was used forde-protection of Boc group to afford the desired amine 86 as a TFA saltin 60% yield.

¹H NMR (DMSO-d₆, 400 MHz): δ 11.44 (s, 1H), 8.87 (d, J=6.4 Hz, 1H),8.68-8.40 (m, 2H), 8.23 (dd, J=11.2, 8.8 Hz, 4H), 7.67 (d, J=8.8 Hz,2H), 7.60 (d, J=8.8 Hz, 2H), 5.36 (s, 2H), 5.17 (s, 2H), 4.86-4.79 (m,1H), 4.07 (d, J=7.6 Hz, 4H).

Step 3 & 4

The similar procedures (coupling & deprotection) with the synthesis ofcarbapenem 63 were used to afford the desired carbapenem 87 in 44% yieldover two-steps except that DIEA was used instead of lutidine base inTHF/toluene mixed solvent (1/10 ratio).

¹H NMR (CDCl₃, 400 MHz): δ 11.67 (s, 1H), 8.48 (d, J=6.8 Hz, 1H),8.21-8.13 (m, 6H), 7.60 (dt, J=6.8, 2.0 Hz, 2H), 7.49 (dt, J=5.2, 2.0Hz, 2H), 7.46 (dt, J=4.8, 2.0 Hz, 2H), 5.43 (d, J=13.6 Hz, 1H), 5.23 (s,2H), 5.16 (d, J=13.6 Hz, 1H), 5.14 (s, 2H), 4.56 (s, J=6.4 Hz, 1H), 4.20(p, J=6.4 Hz, 1H), 4.12 (dd, J=10.0, 3.2 Hz, 1H), 3.83 (d, J=14.4 Hz,1H), 3.58 (m, 1H), 3.22 (d, J=13.2 Hz, 1H), 3.21 (m, 1H), 2.95 (t, J=5.1Hz, 1H), 2.83 (t, J=5.1 Hz, 1H), 1.28 (d, J=6.4 Hz, 3H), 1.09 (d, J=7.2Hz, 3H).

Step 5

According to General Method H, OH compound 87 (200 mg, 0.24 mmol), 5%Pt/C (280 mg) in IPA (5 mL), THF (12 mL), DI water (6 mL) and pH 6buffer (4 mL) were reacted for 0.5 h afford the desired final product 88(30 mg, 37%).

¹H NMR (D₂O, 400 MHz): δ 4.04-3.94 (m, 3H), 3.61 (d, J=12.8 Hz, 1H),3.51 (t, J=7.6 Hz, 1H), 3.46 (t, J=7.6 Hz, 1H), 3.20 (dd, J=6.0, 2.8 Hz,1H), 3.11 (d, J=13.2 Hz, 1H), 2.98 (t, J=7.2 Hz, 1H), 2.93 (dd, J=9.6,7.2 Hz, 1H), 2.85 (d, J=7.2 Hz, 1H), 1.07 (d, J=6.4 Hz, 3H), (d, J=7.6Hz, 3H).

Example 18 Synthesis of Compound 93

Step 1

The similar procedure with side chain, 23, synthesis was used forprotection of nitrogen atom to afford the desired carbamate 89 in 98%yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (d, J=8.8 Hz, 2H), 7.50 (d, J=8.4 Hz,2H), 5.21 (br s, 1H), 5.19 (s, 2H), 4.47 (m, 1H), 4.24 (t, J=9.2 Hz,2H), 3.76 (dd, J=8.8, 1.2 Hz, 2H), 1.45 (s, 9H).

Step 2

The similar procedure with side chain, 61, synthesis was used forde-protection of Boc group to afford the desired amine 90 as a TFA saltin 92% yield.

¹H NMR (DMSO-d6/CDCl₃, 400 MHz): δ 10.25 (br s, 1H), 9.35 (br s, 1H),8.03 (d, J=8.8 Hz, 2H), 7.76 (d, J=8.8 Hz, 1H), 7.35 (d, J=8.8 Hz, 2H),5.02 (s, 2H), 4.53 (s, J=8.4 Hz, 1H), 4.07 (t, J=10.8 Hz, 2H), 3.99 (d,J=10.8 Hz, 2H).

Step 3

According to the general coupling method D, the desired TES compound 91was synthesized in 67% yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (d, J=8.8 Hz, 2H), 8.20 (d, J=8.8 Hz,2H), 7.66 (d, J=8.8 Hz, 2H), 7.49 (d, J=8.4 Hz, 2H), 5.45 (d, J=13.2 Hz,1H), 5.22 (br s, 1H), 5.21 (d, J=13.6 Hz, 1H), 5.17 (s, 2H), 4.34 (s,J=7.6 Hz, 1H), 4.23 (p, J=6.0 Hz, 1H), 4.16 (dd, J=10.4, 3.2 Hz, 1H),3.92 (d, J=10.4 Hz, 1H), 3.67 (t, J=7.2 Hz, 1H), 3.63 (t, J=7.2 Hz, 1H),3.27-3.21 (m, 3H), 2.99 (t, J=6.4 Hz, 1H), 2.88 (t, J=6.4 Hz, 1H), 1.25(d, J=6.0 Hz, 3H), 1.13 (d, J=7.6 Hz, 3H), 0.93 (t, J=8.0 Hz, 9H), 0.58(q, J=8.0 Hz, 6H).

Step 4

According to the general procedure for the removal of TES group (MethodE), the desired OH compound 92 was synthesized in 81% yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (d, J=8.8 Hz, 2H), 8.20 (d, J=8.8 Hz,2H), 7.65 (d, J=8.4 Hz, 2H), 7.48 (d, J=8.4 Hz, 2H), 5.47 (d, J=13.6 Hz,1H), 5.27 (br s, 1H), 5.19 (d, J=14.0 Hz, 1H), 5.17 (s, 2H), 4.33 (s,J=6.8 Hz, 1H), 4.25 (p, J=6.4 Hz, 1H), 4.19 (dd, J=6.0, 3.2 Hz, 1H),3.93 (d, J=14.0 Hz, 1H), 3.67-3.60 (m, 2H), 3.32-3.24 (m, 3H), 3.01 (t,J=6.8 Hz, 1H), 2.92 (t, J=6.8 Hz, 1H), 2.31 (br s, 1H), 1.33 (d, J=6.4Hz, 3H), 1.14 (d, J=7.2 Hz, 3H).

Step 5

According to the general procedure for the removal of PNB groups (MethodG), the desired final product 92 was synthesized in 22% yield.

¹H NMR (D₂O, 400 MHz): δ 4.04-3.95 (m, 2H), 3.87-3.76 (m, 3H), 3.67 (s,J=5.4 Hz, 1H), 3.52 (d, J=12.8 Hz, 1H), 3.37 (t, J=7.6 Hz, 1H), 3.25 (m,1H), 3.22 (dd, J=6.0, 2.8 Hz, 1H), 2.93 (m, 1H), 1.05 (d, J=6.4 Hz, 3H),0.89 (d, J=7.6 Hz, 3H).

Example 19 Synthesis of Compound 99

Step 1

The similar procedure with side chain, 3, synthesis was used for thesubstitution reaction of the mesylate group to afford the desirednitrile compound 93 in 73% yield.

¹H NMR (CDCl₃, 400 MHz): δ 3.78-3.37 (m, 4H), 3.12-3.07 (m, 1H),2.27-2.16 (m, 2H), 1.49 (s, 9H).

Step 2

To a solution of LAH (1.22 g, 32.1 mmol) in ether (15 mL) was added asolution of the nitrile compound 93 (2.5 g, 12.8 mmol) in ether (15 mL)at ice-bath. After overnight at 0° C., it was quenched with 25% NaOH andextracted with ether three times. The extract was dried over anhydrousMgSO₄ and concentrated under a reduced pressure. The crude 94 (1.56 g,61%) was used on the next reaction without further purification.

¹H NMR (CDCl₃, 400 MHz): δ 3.56-3.36 (m, 2H), 3.28 (m, 1H), 2.98 (m,1H), 2.70 (m, 2H), 2.20 (m, 1H), 1.98 (br s, 1H), 1.58 (m, 1H), 1.45 (s,9H), 1.06 (br s, 2H).

Step 3

The similar procedure with side chain, 60, synthesis was used forguanidation to afford the desired guanidine 95 in 82% yield.

¹H NMR (CDCl₃, 400 MHz): δ 11.77 (s, 1H), 8.39 (t, J=5.2 Hz, 1H),8.26-8.20 (m, 4H), 7.55 (d, J=8.4 Hz, 2H), 7.53 (d, J=8.4 Hz, 2H), 5.27(s, 2H), 5.22 (s, 2H), 3.57-3.42 (m, 2H), 3.31 (m, 1H), 3.49 (m, 1H),2.02 (m, 1H), 1.61 (m, 1H), 1.45 (s, 9H).

Step 4

The similar procedure with side chain, 24, synthesis was used forde-protection of Boc group to afford the desired amine 96 as a TFA saltin quantitative yield.

¹H NMR (CDCl₃, 400 MHz): δ 11.71 (br s, 1H), 8.51 (t, J=5.6 Hz, 1H),8.23 (d, J=8.8 Hz, 2H), 8.19 (d, J=8.8 Hz, 2H), 7.53 (d, J=8.4 Hz, 4H),5.27 (s, 2H), 5.20 (s, 2H), 3.62-3.42 (m, 2H), 3.44-3.34 (m, 2H), 3.29(m, 1H), 3.05 (dd, J=11.6, 7.6 Hz, 1H), 2.75 (m, 1H), 2.18 (s, J=7.6 Hz,1H), 1.78 (dq, J=13.6, 8.0 Hz, 1H).

Step 5

According to the general coupling method B, the desired TES compound 97was synthesized in 42% yield.

¹H NMR (CDCl₃, 400 MHz): δ 11.78 (br s, 1H), 8.47 (t, J=4.8 Hz, 1H),8.26-8.19 (m, 6H), 7.67 (d, J=8.8 Hz, 2H), 7.65 (d, J=9.2 Hz, 2H), 7.54(d, J=8.8 Hz, 2H), 5.45 (d, J=14.0 Hz, 1H), 5.32 (s, 2H), 5.27 (S, 2H),5.22 (d, J=14.0 Hz, 1H), 4.26 (p, J=6.0 Hz, 1H), 4.19 (dd, J=10.4, 3.2Hz, 1H), 4.06 (m, 1H), 3.82 (d, J=14.4 Hz, 1H), 3.43 (m, 2H), 3.34 (d,J=14.4 Hz, 1H), 3.30 (m, 1H), 3.22 (dd, J=5.6, 3.2 Hz, 1H), 2.63 (m,1H), 2.63-2.35 (m, 4H), 1.19 (d, J=7.2 Hz, 3H), 1.15 (d, J=6.4 Hz, 3H),0.93 (t, J=7.6 Hz, 92H), 0.59 (q, J=7.6 Hz, 6H).

Step 6

According to the general procedure for the removal of TES group (MethodE), the desired OH compound 98 was synthesized in 77% yield.

¹H NMR (CDCl₃, 400 MHz): δ 11.75 (s, 1H), 8.47 (t, J=4.8 Hz, 1H),8.25-8.19 (m, 6H), 7.65 (d, J=8.8 Hz, 2H), 7.55 (d, J=7.6 Hz, 2H), 7.53(d, J=8.4 Hz, 2H), 5.49 (d, J=14.0 Hz, 1H), 5.27 (s, 2H), 5.22 (s, 2H),5.21 (d, J=13.6 Hz, 1H), 4.26 (d, J=6.0 Hz, 1H), 4.18 (dd, J=10.0, 3.2Hz, 1H), 4.06 (m, 1H), 3.80 (d, J=14.4 Hz, 1H), 3.51-3.20 (m, 3H), 3.35(d, J=14.4 Hz, 1H), 3.25 (dd, J=6.8, 2.8 Hz, 1H), 2.69 (m, 1H),2.50-2.41 (m, 3H), 1.99 (m, 1H), 1.52 (m, 1H), 1.35 (d, J=6.4 Hz, 3H),1.14 (d, J=7.2 Hz, 3H).

Step 7

According to the general procedure for the removal of PNB groups (MethodF), the desired final product 99 was synthesized in 15% yield.

¹H NMR (D₂O, 400 MHz): δ 4.01 (m, 2H), 3.89 (m, 1H), 3.48 (d, J=13.2 Hz,1H), 3.20 (m, 2H), 3.00 (m, 2H), 2.58 (m, 2H), 2.40 (m, 2H), 2.13 (m,1H), 1.60 (m, 1H), 1.07 (br s, 3H), 0.96 (m, 1H), 0.90 (br s, 3H).

Example 20 Synthesis of Compound 106

Step 1

To a slurry of S-methylisothiourea hemisulfate (10 g, 71.9 mmol) andsodium carbonate (35 g, 330 mmol) in DCM (75 mL) was added slowly water(15 mL) at rt and followed by a dropwise addition of methanesulfonylchloride (5.56 mL, 71.8 mmol). After overnight at rt, the liquid wasdecanted and the solid was extracted with DCM. The combined organiclayer was washed with 10% citric acid in water, dried over anhydrousMgSO₄ and concentrated to afford a white solid 100 (6 g, 50%).

¹H NMR (CDCl₃, 400 MHz): δ 3.02 (s, 3H), 2.40 (s, 3H).

Step 2

The similar procedure with side chain, 23, synthesis was used forprotection of nitrogen atom to afford the desired product 101 in 85%yield.

¹H NMR (CDCl₃, 400 MHz): δ 10.45 (s, 1H), 8.24 (d, J=8.8 Hz, 2H), 7.54(d, J=8.8 Hz, 2H), 5.29 (s, 2H), 3.11 (s, 3H), 2.37 (s, 3H).

Step 3

The similar procedure with side chain, 60, synthesis was used forguanidation to afford the desired guanidine 102 in 52% yield.

¹H NMR (CDCl₃, 400 MHz): δ 10.23 (s, 1H), 8.34 (s, 1H), 8.22 (d, J=8.8Hz, 2H), 7.51 (d, J=8.8 Hz, 2H), 5.26 (s, 2H), 4.43 (br s, 1H), 3.62 (m,1H), 3.43 (m, 2H), 3.29-3.20 (m, 1H), 3.00 (s, 3H), 2.16 (m, 1H), 1.87(m, 1H).

Step 4

The similar procedure with side chain, 61, synthesis was used forde-protection of Boc group to afford the desired amine 103 as a TFA saltin quantitative yield.

¹H NMR (CDCl₃, 400 MHz): δ 10.16 (s, 1H), 9.96 (br s, 1H), 9.74 (br s,1H), 8.48 (d, J=6.0 Hz, 1H), 8.21 (d, J=8.8 Hz, 2H), 7.52 (d, J=8.8 Hz,2H), 5.26 (s, 2H), 4.55 (m, 1H), 3.58-3.35 (m, 3H), 3.00 (s, 3H), 2.58(br s, 1H), 2.42 (s, J=6.8 Hz, 1H), 2.07 (m, 1H).

Step 5 & 6

The similar procedures (coupling & deprotection) with the synthesis ofcarbapenem 63 were used to afford the desired OH compound 105 in 56%yield over two-steps.

¹H NMR (CDCl₃, 400 MHz): δ 10.20 (s, 1H), 8.45 (d, J=7.2 Hz, 1H), 8.22(d, J=8.8 Hz, 2H), 8.19 (d, J=8.4 Hz, 2H), 7.63 (d, J=8.8 Hz, 2H), 7.52(d, J=8.8 Hz, 2H), 5.46 (d, J=13.6 Hz, 1H), 5.25 (s, 2H), 5.19 (d,J=14.0 Hz, 1H), 4.39 (m, 1H), 4.25 (m, 1H), 4.22 (dd, J=10.0, 3.2 Hz,1H), 3.84 (d, J=14.8 Hz, 1H), 3.37 (d, J=14.8 Hz, 1H), 3.36 (m, 1H),3.27 (dd, J=6.8, 3.2 Hz, 1H), 2.99 (s, 3H), 2.82 (m, 1H), 2.58-2.51 (m,2H), 2.42 (q, J=6.8 Hz, 1H), 2.43 (m, 2H), 1.68 (m, 1H), 1.34 (d, J=6.0Hz, 3H), 1.18 (d, J=7.2 Hz, 3H).

Step 7

According to the general procedure for the removal of PNB groups (MethodF), the desired final product 106 was synthesized in 33% yield.

¹H NMR (D₂O, 400 MHz): δ 4.30 (br s, 1H), 4.10-4.05 (m, 2H), 3.98 (m,1H), 3.65 (m, 1H), 3.32 (dd, J=5.6, 2.8 Hz, 1H), 3.06 (p, J=6.0 Hz, 1H),3.50-3.00 (m, 4H), 2.87 (s, 3H), 2.37 (br s, 1H), 1.92 (m, 1H), 1.12 (d,J=6.4 Hz, 3H), 1.02 (d, J=7.6 Hz, 3H).

Example 21 Synthesis of Compound 111

Step 1

To a suspended mixture of iso-S-methylthiourea HI salt (5.7 g, 21.9mmol) in DCM (100 mL) was added 220 mL of NaOH (0.1 N) at 0° C. To themixture was added dropwise a solution of p-nitrobenzylchloro formate(4.96 g, 23 mmol) in DCM (20 ml) and 1.0 N NaOH (23 mL) simultaneouslywhile keeping pH above 10. The mixture was warmed up gradually to rtovernight, extracted with DCM and concentrated to afford a white solid106 (2.9 g, 43%).

¹H NMR (CDCl₃, 400 MHz): δ 12.29 (br s, 1H), 8.23 (d, J=8.8 Hz, 2H),7.57 (d, J=8.4 Hz, 2H), 5.28 (s, 2H), 2.42 (s, 3H), 2.21 (s, 3H).

Step 2

The similar procedure with side chain, 60, synthesis was used forguanidation to afford the desired guanidine 107 in 37% yield.

¹H NMR (CDCl₃, 400 MHz): δ 12.21 (br s, 1H), 9.32 (br s, 1H), 8.21 (d,J=8.8 Hz, 2H), 7.58 (d, J=8.8 Hz, 2H), 5.22 (br s, 2H), 4.65 (br s, 1H),3.68 (m, 1H), 3.50-3.25 (m, 3H), 2.28 (s, 3H), 1.90 (m, 1H), 1.42 (s,9H).

Step 3

The similar procedure with side chain, 61, synthesis was used forde-protection of Boc group to afford the desired amine 108 as a TFA saltin 92% yield.

¹H NMR (CDCl₃, 400 MHz): δ 11.96 (s, 1H), 10.45 (br s, 1H), 9.78 (br s,1H), 9.40 (d, J=6.0 Hz, 1H), 8.21 (d, J=8.8 Hz, 2H), 7.52 (d, J=8.8 Hz,2H), 5.20 (s, 2H), 4.60 (br s, 1H), 3.58 (m, 2H), 3.37 (m, 2H), 2.45 (m,1H), 2.20 (s, 3H), 2.12 (m, 1H).

Step 4 & 5

The similar procedures (coupling & deprotection) with the synthesis ofcarbapenem 63 were used to afford the desired OH compound 110 in 29%yield over two-steps.

¹H NMR (CDCl₃, 400 MHz): δ 12.03 (s, 1H), 9.35 7.52 (d, J=8.0 Hz, 1H),8.17 (d, J=8.8 Hz, 4H), 7.61 (d, J=8.4 Hz, 2H), 7.52 (d, J=8.4 Hz, 2H),5.44 (d, J=13.6 Hz, 1H), 5.18 (s, 2H), 5.16 (d, J=13.6 Hz, 1H), 4.59 (m,1H), 4.21 (m, 2H), 3.81 (d, J=14.8 Hz, 1H), 3.36 (d, J=14.8 Hz, 1H),3.35 (m, 1H), 3.26 (dd, J=2.8, 6.4 Hz, 1H), 2.85 (m, 1H), 2.60-2.24 (m,5H), 2.16 (s, 3H), 1.70 (m, 1H), 1.30 (d, J=6.0 Hz, 3H), 1.17 (d, J=7.2Hz, 3H).

Step 6

-   -   to According to the general procedure for the removal of PNB        groups (Method F), the desired final product 111 was synthesized        in 18% yield.

¹H NMR (D₂O, 400 MHz): δ 4.15 (br s, 1H), 4.02 (br s, 2H), 3.69 (br s,1H), 3.59 (br s, 1H), 3.25 (br s, 1H), 3.10-2.70 (m, 4H), 2.28 (br s,1H), 2.05 (m, 1H), 1.95 (s, 3H), 1.84 (br s, 1H), 1.08 (br s, 3H), 0.94(br s, 3H).

Example 22 Synthesis of Compound 115

Step 1

(R)-3-Pyrrolidinecarboxamide 112 was prepared using a similar syntheticmethod as described for the preparation of (S)-3-pyrrolidinecarboxamide9 (Scheme 3 & 4). The amide 112 was obtained from(S)-3-hydroxypyrrolidine HCl salt in 49% yield (five steps).

¹H NMR (CDCl₃, 400 MHz): δ 6.75 (br s, 1H), 6.44 (br s, 1H), 2.98-2.80(m, 4H), 2.70-2.62 (m, 1H), 1.92-1.74 (m, 2H).

Step 2

Compound 113 was achieved in 64% yield using general Method A asdescribed for the Pd coupling reaction.

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (d, J=8.8 Hz, 2H), 7.66 (d, J=8.8 Hz,2H), 6.45 (br s, 1H), 5.54 (br s, 1H), 5.44 (d, J=14.0 Hz, 1H), 5.22 (d,J=14.0 Hz, 1H), 4.27-4.20 (m, 2H), 3.90 (d, J=14.4 Hz, 1H), 3.39 (d,J=14.4 Hz, 1H), 3.31-3.23 (m, 2H), 2.92-2.82 (m, 3H), 2.64-2.60 (m, 1H),2.36-2.29 (m, 1H), 2.22-2.13 (m, 1H), 2.05-1.96 (m, 1H), 1.24 (d, J=6.4Hz, 3H), 1.17 (d, J=7.2 Hz, 3H), 0.93 (t, J=8.0 Hz, 9H), 0.59 (q, J=8.0Hz, 6H).

Step 3

Compound 114 was prepared in 89% yield using general Method E asdescribed for the removal of the TES group.

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (d, J=8.8 Hz, 2H), 7.65 (d, J=8.8 Hz,2H), 6.47 (br s, 1H), 5.55 (br s, 1H), 5.48 (d, J=13.6 Hz, 1H), 5.20 (d,J=13.6 Hz, 1H), 4.28-4.23 (m, 2H), 3.90 (d, J=14.4 Hz, 1H), 3.39 (d,J=14.4 Hz, 1H), 3.37-3.27 (m, 2H), 2.95-2.81 (m, 3H), 2.63-2.59 (m, 1H),2.36-2.30 (m, 1H), 2.22-2.13 (m, 1H), 2.04-1.96 (m, 1H), 1.34 (d, J=6.4Hz, 3H), 1.18 (d, J=7.6 Hz, 3H).

Step 4

The final product 115 was obtained in 76% yield according to a similarprocedure as described for general Method F using THF, IPA and 0.25 Mphosphate buffer solution (pH 7).

¹H NMR (D₂O, 400 MHz): δ 4.05-3.99 (m, 2H), 3.84-3.74 (m, 2H), 3.27-3.25(m, 1H), 3.22-2.97 (m, 6H), 2.23-2.13 (m, 1H), 1.98-189 (m, 1H), 1.06(d, J=6.4 Hz, 3H), 0.95 (d, J=7.2 Hz, 3H).

Example 23 Synthesis of Compound 120

Step 1

To a stirred solution of ester 14 (0.693 g, 2.0 mmol) in THF (20 mL) at0° C. was added 2.0 M solution of Me₂NH in THF dropwise. The resultingsolution was gradually warmed up to room temperature, and stirring wascontinued overnight. The reaction mixture was then filtered, and thefiltrate was evaporated under reduced pressure. The residue was treatedwith DCM and DI water. The aqueous layer was separated and extractedwith DCM. The combined organic layer was washed with brine, dried overNa₂SO₄ and evaporated under reduced pressure. The crude material waspurified by silica gel column chromatography, using DCM/MeOH withgradient as an eluent, to give the desired amide 116 (0.486 g, 88%).

¹H NMR (CDCl₃, 400 MHz): δ 7.30-7.22 (m, 5H), 5.06 (s, 2H), 3.68-3.44(m, 3H), 3.40-3.33 (m, 1H), 3.24-3.13 (m, 1H), 2.99 (s, 3H), 2.89 (s,3H), 2.20-1.96 (m, 2H).

Step 2

Amine 117 was achieved in quantitative yield upon a standardhydrogenolytic Cbz-deprotection.

¹H NMR (CDCl₃, 400 MHz): δ 7.99 (br s, 1H), 3.38-3.14 (m, 5H), 3.02 (s,3H), 2.88 (s, 3H), 2.88 (s, 3H), 2.22-2.09 (m, 1H), 2.03-1.92 (m, 1H).

Step 3

The desired coupling product 118 was obtained in 60% yield using generalMethod A as described for the Pd coupling reaction.

¹H NMR (CDCl₃, 400 MHz): δ 8.20 (d, J=8.4 Hz, 2H), 7.65 (d, J=8.8 Hz,2H), 5.43 (d, J=13.6 Hz, 1H), 5.23 (d, J=13.6 Hz, 1H), 4.26-4.18 (m,2H), 4.11-4.04 (m, 1H), 3.46-3.37 (m, 2H), 3.30-3.21 (m, 2H), 3.08-2.88(m, 3H), 3.02 (s, 3H), 2.93 (s, 3H), 2.84-2.58 (m, 2H), 2.08-1.98 (m,2H), 1.23 (d, J=6.0 Hz, 3H), 1.15 (dd, J=7.2, 2.0 Hz, 3H), 0.93 (t,J=8.0 Hz, 9H), 0.59 (t, J=8, 0 Hz, 6H).

Step 4

The TES group in compound 118 was removed using general Method E toafford the OH-compound 119 in 63% yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (dd, J=6.8, 2.0 Hz, 2H), 7.64 (d, J=8.8Hz, 2H), 5.46 (d, J=14.0 Hz, 1H), 5.21 (d, J=14.0 Hz, 1H), 4.27-4.19 (m,2H), 3.91-3.84 (m, 1H), 3.52-3.31 (m, 2H), 3.25 (dd, J=6.4, 2.8 Hz, 1H),3.21-3.13 (m, 1H), 3.01 (s, 3H), 3.93 (s, 3H), 2.88-2.82 (m, 3H),2.72-2.56 (m, 3H), 2.27 (br s, 1H), 2.10-1.95 (m, 2H), 2.10-1.95 (m,2H), 1.33 (dd, J=6.4, 1.2 Hz, 3H), 1.16 (dd, J=7.6, 3.2 Hz, 3H).

Step 5

The PNB group in compound 119 was removed using general Method F,running hydrogenation in THF, IPA and 0.25 M phosphate buffer solution(pH 7), to afford the desired product 120 in 62% yield.

¹H NMR (D₂O, 400 MHz): δ 4.11-4.02 (m, 2H), 3.81-3.68 (m, 2H), 3.52-3.43(m, 1H), 3.32-3.28 (m, 1H), 3.20-2.97 (m, 5H), 2.93 (s, 3H), 2.76 (s,3H), 2.26-2.15 (m, 1H), 1.92-1.84 (m, 1H), 1.12 (d, J=6.0 Hz, 3H), 0.99(d, J=7.2 Hz, 3H).

Example 24 Synthesis of Compound 127

Step 1

Boc-Protected nitrile 93 (3.93 g, 20.0 mmol) was dissolved in conc. HCl(20 mL). The resulting solution was heated at 100° C. for 3 h. Then thereaction mixture was cooled down and evaporated under reduced pressure.The residue was dried under high vacuum and then dissolved in themixture of acetone (50 mL) and water (50 mL). The resulting solution wascooled to 0° C. and treated slowly with Na₂CO₃ (6.36 g, 60.0 mmol), as asolid, followed by (Boc)₂O (4.80 g, 22.0 mmol). The reaction mixture wasstirred and allowed to warm up rt overnight. Then acetone was removedunder reduced pressure and an aqueous solution was acidified with 6 NHCl to pH 1, and extracted with EtOAc (×4). The combined organic layerswere washed with brine, dried over Na₂SO₄, and concentrated underreduced pressure to give the desired acid 121 (3.49 g, 81%).

¹H NMR (CDCl₃, 400 MHz): δ 9.30 (br s, 1H), 3.66-3.32 (m, 4H), 3.12-3.04(m, 1H), 2.18-2.12 (m, 2H), 1.45 (s, 9H).

Step 2

Compound 122 was prepared in 74% yield using a similar procedure asdescribed for Cbz-protected analog 14 (Scheme 3).

¹H NMR (CDCl₃, 400 MHz): δ 3.73-3.32 (m, 5H), 2.84 (s, 4H), 2.32-2.26(m, 2H), 1.45 (s, 9H).

Step 3

Amide 123 was synthesized in quantitative yield according to proceduredescribed in Scheme 23 for preparation of compound 116.

¹H NMR (CDCl₃, 400 MHz): δ 6.51 (br s, 1H), 3.57-3.36 (m, 3H), 3.27-3.20(m. 1H), 2.87-2.79 (m, 1H), 2.73 (d, J=4.8 Hz, 3H), 2.12-1.97 (m. 2H),1.38 (s, 9H).

Step 4

Compound 123 (0.685 g, 3.0 mmol) was treated with cold 4 M HCl indioxane (10.0 mL). The reaction mixture was stirred at 0° C. for 3 h(monitoring by TLC). The HCl salt of the desired amine 124 graduallyprecipitated from the reaction mixture as a white solid which wasfiltered off, washed with fresh dioxane followed by diethyl ether anddried (0.384 g, quantitative yield).

¹H NMR (DMSO-d₆, 400 MHz): δ 9.48 (br s, 1H), 9.20 (br s, 1H), 8.20 (d,J=4.0 Hz, 1H), 3.31-3.23 (m, 1H), 3.18-3.07 (m, 2H), 2.99 (q, J=7.6 Hz,1H), 2.57 (d, J=4.8 Hz, 3H), 2.13-2.04 (m, 1H), 1.92-1.83 (m, 1H).

Step 5

Compound 125 was synthesized in 29% yield by the Pd coupling reaction ofCPI intermediate with free amine 124 using general Method A.

¹H NMR (CDCl₃, 400 MHz): δ 8.19 (dd, J=7.2, 2.0 Hz, 2H), 7.65 (d, J=8.8Hz, 2H), 6.50 (d, J=2.0 Hz, 1H), 5.43 (d, J=14.0 Hz, 1H), 5.20 (d,J=14.0 Hz, 1H), 4.26-4.17 (m, 2H), 3.93 (d, J=14.0 Hz, 1H), 3.35 (d,J=14.4 Hz, 1H), 3.28-3.22 (m, 2H), 2.89-2.78 (m, 3H), 2.75 (d, J=4.8 Hz,3H), 2.53-2.47 (m, 2H), 2.16-2.07 (m, 1H), 2.02-1.93 (m, 1H), 1.24 (d,J=7.2 Hz, 3H), 1.17 (d, J=7.2 Hz, 2H), 0.92 (t, J=8.0 Hz, 9H), 0.60 (q,J=8.0 Hz, 6H).

Step 6

Using general Method E, the TES group was removed from compound 125 toafford the OH-compound 126 in 81% yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.20 (d, J=8.8 Hz, 2H), 7.63 (d, J=8.8 Hz,2H), 6.40 (d, 4.8 Hz, 1H), 5.46 (d, J=14.0 Hz, 1H), 5.19 (d, J=14.0 Hz,1H), 4.27-4.20 (m, 2H), 3.89 (d, J=14.8 Hz, 1H), 3.34-3.26 (m, 3H),2.87-2.77 (m, 3H), 2.75 (d, J=4.8 Hz, 3H), 2.55 (br s, 1H), 2.49-2 43(m, 2H), 2.15-2.05 (m, 1H), 2.00-1.92 (m, 1H), 1.33 (d, J=6.4 Hz, 3H),1.18 (d, J=7.2 Hz, 3H).

Step 7

The final product 127 was obtained in 69% yield by a similar procedureas described for general Method F using THF, IPA and 0.25 M phosphatebuffer pH 7.0.

¹H NMR (D₂O, 400 MHz): δ 4.09-4.04 (m, 2H), 3.83-3.73 (m, 2H), 3.31-2.98(m, 6H), 2.56 (s, 3H), 2.20-2.10 (m, 1H), 1.98-1.90 (m, 1H), 1.11 (d,J=6.4 Hz, 3H), 0.99 (d, J=7.2 Hz, 3H).

Example 25 Synthesis of Compound 132

Step 1

To a solution of acid 121 (0.646 g, 3.0 mmol) in anhydrous DCM (50 mL),a 1.0 M solution of DCC in DCM (4.5 mL, 0.928 g, 4.5 mmol) was added,followed by methanesulfonamide (0.285 g, 3.0 mmol) and DMAP (0.366 g 3.0mmol). The reaction mixture was stirred at room temperature overnight.Then resulting precipitate was removed by filtration. The filtrate wasevaporated under reduced pressure to dryness. The residue was purifiedby silica gel flash chromatography to give the desired product 128 in52% yield.

¹H NMR (CDCl₃, 400 MHz): δ 3.68-3.44 (m, 3H), 3.38-3.31 (m, 1H), 3.27(s, 3H), 3.11-3.03 (m, 1H), 2.25-2.08 (m, 1H), 1.43 (s, 9H).

Step 2

Upon a standard Boc-deprotection procedure with TFA, as describedearlier, TFA salt 129 of the desired amine was prepared in 92% yield andused for the next step.

¹H NMR (DMSO-d₆, 400 MHz): δ 8.88 (br s, 2H), 3.35-3.30 (m, 2H), 3.25(s, 3H), 3.22-3.13 (m, 3H), 2.24-2.15 (m, 1H), 2.02-1.93 (m, 1H).

Step 3

General Method B for the Pd coupling reaction gave the desiredTES-protected product 130 in 48% yield together with TES-deprotectedproduct 131 (21% yield).

Product 130: ¹H NMR (CDCl₃, 400 MHz): δ 8.19 (d, J=8.8 Hz, 2H), 7.65 (d,J=8.8 Hz, 2H), 5.42 (d, J=14.0 Hz, 1H), 5.25 (d, J=14.0 Hz, 1H), 4.58(d, J=13.2 Hz, 1H), 4.32-4.22 (m, 2H), 3.73 (d, J=13.6 Hz, 1H),3.45-3.36 (m, 3H), 3.29-3.22 (m, 3H), 3.14 (s, 3H), 3.14-3.08 (m, 1H),2.42-2.32 (m, 1H), 2.26-2.18 (m, 1H), 1.20 (d, J=6.0 Hz, 3H), 1.15 (d,J=7.2 Hz, 3H), 0.90 (t, J=8.0 Hz, 9H), 0.56 (q, J=8.0 Hz, 6H).

Step 4

The TES product was deprotected using general Method E to give compound131 in 93% yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.15 (d, J=8.8 Hz, 2H), 7.60 (d, J=8.8 Hz,2H), 5.40 (d, J=14.0 Hz, 1H), 5.19 (d, J=13.6 Hz, 1H), 4.30 (d, J=13.6Hz, 1H), 4.22 (dd, J=10.0, 2.8 Hz, 1H), 4.10 (q, J=6.4 Hz, 1H), 3.59 (d,J=13.6 Hz, 1H), 3.37-3.24 (m, 3H), 3.21 (dd, J=6.8, 2.8 Hz, 1H),3.08-2.96 (m, 1H), 3.04 (s, 3H), 2.86-2.78 (m, 2H), 2.30-2.21 (m, 1H),2.13-2.04 (m, 1H), 1.25 (d, J=6.4 Hz, 3H), 1.11 (d, J=7.6 Hz, 3H).

Step 5

The final product 132 was obtained using general Method F in 68% yield.

¹H NMR (D₂O, 400 MHz): δ 4.08-4.02 (m, 2H), 3.92 (s, 2H), 3.70-3.36 (m,2H), 3.30 (dd, J=6.0, 2.8 Hz, 1H), 3.26-3.10 (m, 3H), 3.08-2.98 (m, 3H),2.84 (s, 3H), 2.30-1.94 (m, 2H), 1.08 (d, J=6.4 Hz, 3H), 0.98 (d, J=7.2Hz, 3H).

Example 26 Synthesis of Compound 137

Step 1

Compound 13 (0.400 g, 1.6 mmol) was dissolved in anhydrous ACN (20 mL)and cooled to 0° C. To the resulting solution glycinamide hydrochloride(0.230 g, 2.08 mmol), EDCI×HCl (0.461 g. 2.4 mmol), HOBt (0.324 g, 2.4mmol) and DIEA (0.836 mL, 4.8 mmol) were added, and the reaction mixturewas stirred under N₂ atmosphere at room temperature for 24 h. Then thesolution was evaporated under reduced pressure. The residue wasdissolved in EtOAc and washed with 1 M HCl and brine. The organic layerwas dried over Na₂SO₄ and concentrated. The crude product was purifiedby silica gel flash chromatography eluting with DCM/MeOH with gradientto afford the desired amide 133 (0.480 g, 98%).

¹H NMR (CDCl₃, 400 MHz): δ 7.61 and 7.45 (t+t, J=5.2 Hz, 1H), 7.30-7.24(m, 5H), 7.01 and 6.96 (br s+br s, 1H), 6.61 and 6.54 (br s+br s, 1H),5.06 (d, J=3.2 Hz, 2H), 3.93-3.76 (m, 2H), 3.66-3.44 (m, 3H), 3.37-3.29(m, 1H), 3.00-2.91 (m, 1H), 2.12-2.01 (m, 2H).

Step 2

Upon a standard hydrogenolytic Cbz-deprotection the corresponding amine134 was achieved in quantitative yield.

¹H NMR (DMSO-d₆, 400 MHz): δ 9.11 (br s, 1H), 8.39 (t, J=1.6 Hz, 1H),7.36 (s, 1H), 7.03 (s, 1H), 3.68-3.57 (m, 2H), 3.31-3.26 (m, 1H),3.21-3.07 (m, 4H), 2.15-2.06 (m, 1H), 1.97-1.88 (m, 1H).

Step 3

The desired coupling product 135 was prepared in 28% yield using generalMethod A.

¹H NMR (CDCl₃, 400 MHz): δ 8.20 (dd, J=6.8, 2.0 Hz, 2H), 7.66 (dd, J=8.8Hz, 2H), 7.32 (br s, 1H), 6.37 (br s, 1H), 5.64 (br s, 1H), 5.44 (d,J=14.0 Hz, 1H), 5.22 (d, J=14.0 Hz, 1H), 4.27-4.20 (m, 2H), 3.94-3.81(m, 3H), 3.42-3.31 (m, 2H), 3.25-3.23 (m, 1H), 2.92-2.83 (m, 3H),2.27-2.12 (m, 2H), 2.03-1.94 (m, 2H), 1.24 (d, J=6.0 Hz, 3H), 1.17 (d,J=7.2 Hz, 3H), 0.93 (t, J=8.0 Hz, 9H), 0.59 (q, J=8.0 Hz, 6H).

Step 4

The TES compound 135 was deprotected using general Method E to givecompound 136 in 61% yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.22 (dd, J=7.6, 2.0 Hz, 2H), 7.70 and 7.41(t+t, J=4.8 Hz, 1H), 7.65 (d, J=8.8 Hz, 2H), 6.70 and 6.61 (br s+br s,1H), 6.57 and 6.40 (br s+br s, 1H), 5.47 (dd, J=13.6, 3.6 Hz, 1H), 5.22(dd, J=13.6, 2.4 Hz, 1H), 4.25-4.19 (m, 2H), 4.05-3.73 (br m, 3H),3.61-3.48 (m, 1H), 3.38 (t, J=14.4 Hz, 1H), 3.25 (dd, J=7.2, 2.8 Hz,1H), 3.03-2.85 (m, 3H), 2.32-2.15 (m, 2H), 2.01-1.91 (m, 2H), 1.36 (dd,J=9.6, 6.0 HZ, 3H), 1.17 (dd, J=7.6, 2.8 Hz, 3H).

Step 5

The final desired product 137 was obtained in 55% yield using a similarprocedure as described for general Method F.

¹H NMR (D₂O, 400 MHz): δ 4.10-4.04 (m, 2H), 3.86-3.80 (m, 2H), 3.75 (s,2H), 3.34-3.30 (m, 2H), 3.24-3.01 (m, 5H), 2.28-2.18 (m, 1H), 2.06-1.97(m, 1H), 1.12 (d, J=5.6 Hz, 3H), 1.00 (d, J=6.4 Hz, 3H).

Example 27 Synthesis of Compound 143

Step 1

1,4-Dioxane (10 mL) was added to a solution of guanidine hydrochloride(0.96 g, 10.0 mmol) and NaOH (0.80 g, 20.0 mmol) in H₂O (10 mL), and theresulting mixture was cooled to 0° C. Next, a solution of 4-nitrobenzylchloroformate (1.66 g, 7.7 mmol) in 1,4-dioxane (15 mL) was slowly addedat 0-5° C. under vigorous stirring. After stirring for an additional 10h at room temperature, the mixture was concentrated under reducedpressure to one-third its original volume and extracted with EtOAc threetimes. The combined extracts were washed with brine and dried overNa₂SO₄. After filtering and a removal of the solvent under reducedpressure, the pure mono-protected guanidine 138 (1.56 g, 85%) wasobtained.

¹H NMR (DMSO-d₆, 400 MHz): δ 8.18 (d, J=2.0 Hz, 2H), 7.57 (d, J=2.0 Hz,2H), 5.69 (br s, 4H), 4.62 (s, 2H).

Step 2

To a cold solution of acid 121 (0.646 g, 3.0 mmol) in anhydrous DCM (60mL), guanidine 138 (0.929 g, 3.9 mmol), EDCI×HCl (0.863 g, 4.5 mmol) andDMAP (0.586 g, 4.8 mmol) were added. The reaction mixture was stirredunder N₂ atmosphere and allowed to warm up. After 24 h the solvent wasevaporated under reduced pressure. The residue was dissolved in EtOAcand washed with 1 N HCl and brine. The organic layer was dried overNa₂SO₄ and evaporated under reduced pressure. The residue was purifiedby silica gel flash chromatography to give the desired product 139 in70% yield.

¹H NMR (DMSO-d₆, 400 MHz): δ 10.15 (br s, 1H), 8.40 (br s. 1H), 8.22 (d,J=8.8 Hz, 2H), 7.62 (d, J=8.8 Hz, 2H), 5.74 (br s, 1H), 5.25 (s, 2H),3.49-3.38 (m, 2H), 3.30-3.21 (m, 2H), 3.30-3.21 (m, 2H), 3.14-3.07 (m,1H), 2.15-1.97 (m, 2H), 1.40 (s, 9H).

Step 3

Compound 140 was achieved in 91% yield upon a standard TFA deprotection.

¹H NMR (CDCl₃, 400 MHz): δ 9.95 (br s, 2H), 8.22 (d, J=6.8 Hz, 2H), 7.51(d, J=6.8 Hz, 2H), 5.24 (s, 2H), 3.60-3.51 (m, 2H), 3.38-3.30 (m, 3H),2.41-2.25 (m, 2H).

Step 4

General Method B for the Pd coupling reaction gave a mixture ofTES-protected product 141 (26%) together with TES-deprotected product142 (40%).

Product 141: ¹H NMR (CDCl₃, 400 MHz): δ 11.50 (br s, 1H), 10.10 (br s,1H), 8.40 (br s, 1H), 8.20-8.16 (m, 4H), 7.63 (d, J=8.8 Hz, 2H), 7.51(d, J=8.8 Hz, 2H), 5.45 (d, J=13.6 Hz, 1H), 5.24 (s, 2H), 5.20 (d,J=13.6 Hz, 1H), 4.77-4.71 (m, 1H), 4.34-4.19 (m, 3H), 3.88-3.83 (m, 2H),3.56-3.28 (m, 5H), 2.49-2.39 (m, 1H), 2.36-2.24 (m, 1H), 1.28 (d, J=6.4Hz, 3H), 1.14 (d, J=7.2 Hz, 3H), 0.91 (t, J=8.0 Hz, 9H), 0.57 (q, J=8.0Hz, 6H).

Step 5

The TES product was deprotected using general Method E to give anadditional amount of compound 142 in 94% yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.60 (d, J=3.2 Hz, 1H), 8.22-8.17 (m, 4H),7.61 (d, J=8.4 Hz, 2H), 7.52 (d, J=8.8 Hz, 2H), 5.44 (d, J=13.6 Hz, 1H),5.25 (s, 2H), 5.17 (d, J=14.0 Hz, 1H), 4.77-4.74 (m, 1H), 4.35 (dd,J=10.4, 3.2 Hz, 1H), 4.27-4.22 (m, 1H), 3.88 (d, J=13.2 Hz, 1H),3.55-3.38 (m, 4H), 3.32 (dd, J=5.2, 2.8 Hz, 1H), 3.20-3.10 (m, 1H),2.50-2.41 (m, 1H), 2.34-2.25 (m, 1H), 2.34-2.25 (m, 1H), 1.29 (d, J=6.4Hz, 3H), 1.15 (d, J=7.2 Hz, 3H).

Step 6

The final product 143 was obtained in 61% yield using a similarprocedure as described for general Method F.

¹H NMR (D₂O, 400 MHz): δ 4.04-4.01 (m, 2H), 3.90 (br s, 2H), 3.29-3.27(m, 2H), 3.23-3.21 (m, 1H), 3.18-3.11 (m, 1H), 3.05-2.95 (m, 3H),2.22-2.10 (m, 1H), 2.04-1.93 (m, 1H), 1.06 (d, J=6.4 Hz, 3H), 0.96 (d,J=7.2 Hz, 3H).

Example 28 Synthesis of Compound 146

Step 1

Compound 144 was synthesized in 82% yield by using general Method A asdescribed for the Pd coupling reaction.

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (dd, J=6.8, 2.0 Hz, 2H), 7.66 (d, J=8.8Hz, 2H), 5.45 (d, J=14.0 Hz, 1H), 5.21 (d, J=14.0 Hz, 1H), 4.33-4.30 (m,1H), 4.24 (t, J=6.0 Hz, 1H), 4.19 (dd, J=10.4, 3.2 Hz, 1H), 3.86 (d,J=14.4 Hz, 1H), 3.38-3.33 (m, 2H), 3.26 (dd, J=5.6, 2.8 Hz, 1H),2.90-2.87 (m, 1H), 2.66 (d, J=10.0 Hz, 1H), 2.44-2.34 (m, 2H), 2.21-2.13(m, 1H), 1.77-1.71 (m, 1H), 1.25 (d, J=6.4 Hz, 3H), 1.17 (d, J=7.6 Hz,3H), 0.93 (t, J=7.6 Hz, 9H), 0.59 (q, J=7.6 Hz, 6H).

Step 2

The TES group in compound 144 was removed using general Method E toafford product 145 in 54% yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (dd, J=6.8, 2.0 Hz, 2H), 7.64 (d, J=8.8Hz, 2H), 5.47 (d, J=14.0 Hz, 1H), 5.20 (d, J=14.0 Hz, 1H0, 4.32-4.29 (m,1H), 4.28-4.20 (m, 1H), 3.85 (d, J=14.4 Hz, 1H), 3.43-3.37 (m, 1H), 3.35(d, J=14.4 Hz, 1H), 3.26 (dd, J=6.0, 2.8 Hz, 1H), 2.92-2.87 (m, 1H),2.66 (d, J=6.0 Hz, 1H), 2.45 (br s, 1H), 2.39-2.30 (m, 2H), 2.21-2.12(m, 1H), 1.77-1.70 (m, 1H), 1.31 (d, J=6.0 Hz, 3H), 1.16 (d, J=7.2 Hz,3H).

Step 3

The final product 146 was obtained in 64% yield using a similarprocedure as described for general Method F.

¹H NMR (D₂O, 400 MHz): δ 4.44 (br s, 1H), 4.11-4.05 (m, 2H), 3.91-3.81(m, 2H), 3.32-3.29 (m, 2H), 3.14-3.03 (m, 4H), 2.20-2.09 (m, 1H),1.89-1.82 (m, 1H), 1.12 (d, J=6.0 Hz, 3H), 1.00 (d, J=6.8 Hz, 3H).

Example 29 Synthesis of Compound 151

Steps 1

According to the procedures described in Scheme 6 for the preparation of(S)-3-aminopyrrolidine 22, (S)-3-hydroxypyrrolidine hydrochloride wasconverted to (R)-3-aminopyrrolidine 147 in 91% yield (four steps).

¹H NMR (CDCl₃, 400 MHz): δ 3.58-3.31 (m, 4H), 3.07-2.97 (m, 1H),2.07-1.99 (m, 1H), 1.68-1.60 (m, 1H), 1.45 (s, 9H).

Step 2

Amine 147 was protected with the PNB group to give compound 148 in 79%yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (d, J=8.8 Hz, 2H), 7.50 (d, J=8.8 Hz,2H), 5.18 (s, 2H), 5.03 (br s, 1H), 4.25-4.22 (m, 1H), 3.60 (dd, J=11.2,6.0 Hz, 1H), 3.43-3.39 (m, 2H), 3.27-3.17 (m, 1H), 2.18-2.10 (m, 1H),1.90-1.78 (m, 1H), 1.44 (s, 9H).

Step 3

A standard Boc-deprotection procedure afforded pyrrolidine 149 in 87%yield.

¹H NMR (DMSO-d₆, 400 MHz): δ 8.80 (br s, 1H), 8.22 (d, J=8.4 Hz, 2H),7.82 (d, J=6.0 Hz, 1H), 7.60 (d, J=8.4 Hz, 2H), 5.17 (s, 2H), 4.15-4.11(m, 1H), 3.34-3.14 (m, 4H), 3.02 (dd, J=11.6, 4.8 Hz, 1H), 2.12-2.02 (m,1H), 1.87-1.79 (m, 1H).

Step 4

-   -   to General Method B for the Pd coupling reaction gave de-TES        compound 150 as a major product (54% yield), together with a        small amount of TES-protected product.

¹H NMR (CDCl₃, 400 Hz): δ 8.13-8.09 (m, 4H), 7.56 (d, J=8.8 Hz, 2H),7.43 (d, J=8.8 Hz, 2H), 5.40 (d, J=13.6 Hz, 1H), 5.13 (d, J=13.6 Hz,1H), 5.11 (s, 2H), 4.60 (d, J=12.8 Hz, 1H), 4.48 (br s, 1H), 4.30 (d,J=10.4 Hz, 1H), 4.22-4.16 (m, 1H), 3.76 (d, J=13.2 Hz, 1H), 3.54-3.48(m, 3H), 3.31-3.18 (m, 3H), 3.06-2.94 (m, 1H), 2.48-2.38 (m, 1H),2.06-1.97 (m, 1H), 1.25 (d, J=6.4 Hz, 3H), 1.11 (d, J=6.8 Hz, 3H).

Step 5

The final product 151 was obtained in 29% yield using general Method F.

¹H NMR (D₂O, 400 MHz): δ 4.09-4.03 (m, 2H), 3.64 (d, J=12.8 Hz, 2H),3.41 (d, J=14.4 Hz, 1H), 3.08-3.02 (m, 2H), 2.80-2.70 (m, 2H), 2.47-2.42(m, 1H), 2.23-2.13 (m, 1H), 1.71-1.64 (m, 1H), 1.12 (d, J=6.4 Hz, 3H),0.96 (d, J=7.2 Hz, 3H).

Example 30 Synthesis of Compound 156

Synthetic methodology and procedures shown above are similar to thosepresented in Scheme 13 for the preparation of carbapenem 64.

Step 1

The reaction of (R)-1-Boc-3-aminopyrrolidine 147 with PNB-protectedS-methylisothiourea 59 afforded guanidine 152 in 93% yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.42 (br s, 1H), 8.25-8.19 (m, 4H), 7.54 (d,J=8.0 Hz, 2H), 7.52 (d, J=8.0 Hz, 2H), 5.26 (s, 2H), 5.22 (s, 2H),4.68-4.62 (m, 1H), 3.70-3.65 (m, 1H), 3.48-3.40 (m, 2H), 3.33-3.20 (m,1H), 2.24-2.16 (m, 1H), 1.93-1.85 (m, 1H), 1.45 (s, 9H).

Step 2

A standard procedure for Boc-deprotection, with TFA afforded the desiredamine 153 isolated as a TFA salt in 98% yield.

¹H NMR (DMSO-d₆, 400 MHz): δ 11.45 (br s, 1H), 8.80 (br s, 2H), 8.43 (d,J=7.2 Hz, 1H), 8.25-8.21 (m, 4H), 7.67 (d, J=8.8 Hz, 2H), 7.60 (d, J=8.8Hz, 2H), 5.35 (s, 2H), 5.19 (s, 2H), 4.69-4.61 (m, 1H), 3.40-3.27 (m,2H), 3.18-3.12 (m, 2H), 2.26-2.17 (m, 1H), 1.95-1.86 (m, 1H).

Step 3

General Method B for the Pd coupling reaction afforded a mixture ofTES-protected product 154 (28% yield), together with de-TES product 155(46% yield).

Product 154: ¹H NMR (CDCl₃, 400 MHz): δ 11.75 (s, 1H), 8.57 (d, J=7.6Hz, 1H), 8.25-8.18 (m, 6H), 7.65 (d, J=8.4 Hz, 2H), 7.56-7.49 (m, 4H),5.45 (d, J=13.6 Hz, 1H), 5.26-5.20 (m, 5H), 4.63 (br s, 1H), 4.29-4.19(m, 2H), 3.86 (d, J=14.4 Hz, 1H), 3.41-3.32 (m, 2H), 3.25 (dd, J=6.4,2.8 Hz, 1H), 3.09 (dd, J=6.8, 2.8 Hz, 1H), 2.85-2.74 (m, 2H), 2.58-2.50(m, 1H), 2.32-2.28 (m, 2H), 1.26 (d, J=6.0 Hz, 3H), 1.18 (d, J=7.6 Hz,3H), 0.95 (t, J=7.6 Hz, 9H), 0.60 (q, J=7.6 Hz, 6H).

Step 4

The TES product was deprotected using general Method E to obtain the OHproduct 155 in 94% yield.

¹H NMR (CDCl₃, 400 MHz): δ 11.74 (s, 1H), 8.62 (br s, 1H), 8.22-8.16 (m,6H), 7.61 (d, J=8.4 Hz, 2H) 7.52 (d, J=8.4 Hz, 2H), 7.51 (d, J=8.8 Hz,2H), 5.43 (d, J=14.0 Hz, 1H), 5.25 (s, 2H), 5.19 (s, 2H), 5.18 (d,J=14.0 Hz, 1H), 4.88 (br s, 1H), 4.69 (d, J=13.2 Hz, 1H), 4.34 (dd,J=10.4, 2.8 Hz, 1H), 4.24 (q, J=6.0 Hz, 1H), 3.86 (d, J=13.2 Hz, 1H),3.76-3.64 (m, 1H), 3.58-3.44 (m, 2H), 3.33-3.20 (m, 2H), 3.10-2.80 (m,2H), 2.60-2.52 (m, 1H), 2.19-2.10 (m, 1H), 1.29 (d, J=6.0 Hz, 3H), 1.14(d, J=7.2 Hz, 3H).

Step 5

The desired final product 156 was achieved in 35% yield using generalMethod F.

¹H NMR (D₂O, 400 MHz): δ 4.04-3.96 (m, 2H), 3.92-3.86 (m, 1H), 3.54 (d,J=13.6 Hz, 1H), 3.25-3.20 (m, 2H), 3.03-2.99 (m, 1H), 2.87-2.83 (m, 1H),2.68-2.60 (m, 1H), 2.51-2.44 (m, 1H), 2.40-2.34 (m, 1H), 2.16-2.11 (m,1H), 1.62-1.57 (m, 1H), 1.06 (d, J=6.4 Hz, 3H), 0.89 (d, J=7.2 Hz, 3H).

Example 31 Synthesis of Compound 163

Step 1

To a cold solution of (S)-1-Boc-3-aminopyrrolidine (6.15 g, 33.0 mmol)in anhydrous DCM (60 mL) was added DIEA (7.5 mL, 42.9 mmol) under N₂atmosphere at 0° C. and followed by addition of bromoacetyl chloride(3.3 mL, 39.6 mmol) dropwise. The reaction mixture was allowed to warmup, and stirring was continued at room temperature for 24 h. Then, themixture was diluted with EtOAc, washed with aqueous NaHCO₃ and brine,dried over Na₂SO₄, filtered, and evaporated under reduced pressure togive a crude material which was purified by flash chromatography onsilica gel eluting with hexane/EtOAc with gradient. The desired product157 was obtained in 80% yield (8.1 g).

¹H NMR (CDCl₃, 400 MHz): δ 6.62 (br s, 1H), 4.48-4.41 (m, 1H), 4.04 (s,1H), 3.86 (s, 1H), 3.66-3.61 (m, 1H), 3.46-3.41 (m, 2H), 3.28-3.17 (m,1H), 2.22-2.12 (m, 1H), 1.93-1.82 (m, 1H), 1.45 (s, 9H).

Step 2

To a solution of compound 157 (6.0 g, 19.53 mmol) in MeOH (110 mL) wasadded NaI (8.78 g, 58.6 mmol) and 28% aqueous NH₄OH (110 mL). Theresulting mixture was stirred at room temperature for 48 h. The mixturewas evaporated under reduced pressure to dryness. The residue waspurified by silica gel flash chromatography eluting with ACN to give 4.6g of the desired product 158 (96.8%).

¹H NMR (CDCl₃, 400 MHz): δ 8.38 and 8.28 (s+s, 1H), 7.64 (br s, 2H),4.51-4.42 (m, 1H), 4.25-4.00 (m, 2H), 3.66-3.34 (m, 4H), 2.20-2.00 (m,2H), 1.41 (s, 9H).

Step 3

To a solution of compound 158 (0.730 g, 3.0 mmol) in dioxane (20 mL) wasadded sulfamide (0.577 g, 6.0 mmol). The resulting mixture was stirredunder reflux. Progress of the reaction was monitored by TLC. Whenreaction was complete the mixture was cooled down and insoluble materialwas filtered off. The filtrate was concentrated under reduced pressure.The residue was treated with EtOAc, and insoluble material was filteredoff again. The filtrate was evaporated under reduced pressure. Theresidue was purified by silica gel flash chromatography using DCM/MeOHwith gradient as an eluent. The desired product 159 was obtained in 56%yield (0.542 g).

¹H NMR (CDCl₃, 400 MHz): δ 7.40 (br s, 1H), 6.08 (br s, 1H), 5.79 (br s,2H), 4.45-4.35 (m, 1H), 3.78-3.20 (m, 6H), 2.15-2.04 (m, 1H), 1.92-1.85(m, 1H), 1.44 (s, 9H).

Step 4

A standard procedure for Boc-deprotection, with 4 M HCl in dioxanefollowed by liberation of free amine with base, afforded the desiredamine 160 in 89% yield.

HCl salt of 160: ¹H NMR (DMSO-d₆, 400 MHz): δ 9.24 (br s, 3H), 8.20 (d,J=2.8 Hz, 1H), 6.70 (br s, 2H), 4.36-4.30 (m, 1H), 3.32-3.23 (m, 2H),3.20-3.14 (m, 1H), 3.04-2.98 (m, 1H), 2.13-2.04 (m, 1H), 1.87-1.79 (m,1H).

Step 5

General Method B for the Pd coupling reaction afforded the desiredproduct 161 in 41% yield.

¹H NMR (CDCl₃, 400 MHz): δ 8.21 (d, J=8, 8 Hz, 2H), 7.66 (d, J=8.4 Hz,2H), 6.91 (d, J=7.6 Hz, 1H), 5.81 (br s, 2H), 5.50 (br s, 1H), 5.43 (d,J=14.0 Hz, 1H), 5.23 (d, J=14.0 Hz, 1H), 4.44-4.37 (m, 1H), 4.27-4.21(m, 2H), 3.88 (d, J=14.4 Hz, 1H), 3.74 (s, 2H), 3.34-3.28 (m, 2H), 3.24(dd, J=4.8, 2.8 Hz, 1H), 2.81-2.75 (m, 1H), 2.69-2.65 (m, 1H), 2.57-2.54(m, 1H), 2.51-2.44 (m, 1H), 2.28-2.19 (m, 1H), 1.70-1.62 (m, 1H), 1.24(d, J=6.4 Hz, 3H), 1.16 (d, J=7.2 Hz, 3H), 0.93 (t, J=8.0 Hz, 9H), 0.59(q, J=8.0 Hz, 6H).

Step 6

Compound 161 (0.32 g, 0.46 mmol) was dissolved in THF (15 mL) and IPA (4mL), and cooled to 0° C. Then, 0.06 N HCl solution was added graduallyto maintain pH value 2.4. The resulting mixture was aged at 0° C. Afterreaction completion, the reaction mixture was neutralized with 0.25 Msodium phosphate buffer pH 7.0 (10 mL), stirred for 10 minutes andtreated with EtOAc. After separation, the aqueous layer was extractedwith EtOAc twice. The combined organic layer was dried over Na₂SO₄ andevaporated under reduced pressure. The residue was purified by flashchromatography on silica gel eluting with DCM/MeOH with gradient to givethe desired OH compound 162 in 60% yield.

¹H NMR (CDCL₃, 400 MHz): δ 8.21 (d, J=8.8 Hz, 2H), 7.63 (d, J=8.8 Hz,2H), 7.20 (d, J=3.6 Hz, 1H), 6.01 (br s, 2H), 5.44 (d, J=13.6 Hz, 1H),5.22 (d, J=13.6 Hz, 1H), 4.36-4.29 (m, 1H), 4.27-4.19 (m, 2H), 3.82 (d,J=14.4 Hz, 1H), 3.69 (s, 2H), 3.53-3.29 (m, 2H), 3.22 (dd, J=6.8, 2.8Hz, 1H), 3.03-3.00 (m, 1H), 2.67 (d, J=9.6 Hz, 1H), 2.29-2.19 (m, 2H),1.73-1.64 (m, 1H), 1.30 (d, J=6.0 Hz, 3H), 1.14 (d, J=7.2 Hz, 3H).

Step 7

General Method F for the removal of the PNB protecting group affordedthe desired final product 163 in 64% yield.

¹H NMR (D₂O, 400 MHz): δ 4.30-4.23 (m, 1H), 4.07-3.99 (m, 2H), 3.73-3.63(m, 2H), 3.57 (s, 2H), 3.26 (dd, J=6.0, 2.8 Hz, 1H), 3.06-2.98 (m, 3H),2.92-2.83 (m, 2H), 2.26-2.17 (m, 1H), 1.81-1.73 (m, 1H), 1.08 (d, J=6.4Hz, 3H), 0.95 (d, J=7.2 Hz, 3H).

Example 32 Synthesis of Compound 167

Step 1

A standard guanidation reaction with 59 afforded the desired product 164in 52% yield.

¹H NMR (DMSO-d₆, 400 MHz): δ 11.50 (br s, 1H), 8.94 (t, J=4.4 Hz, 1H),8.23-8.12 (m, 4H), 7.54-7.50 (m, 4H), 6.68 and 6.62 (br s+br s, 1H),5.28 (s, 2H), 5.18 (s, 2H), 4.44-4.40 (m, 1H), 4.06 (d, J=4.8 Hz, 2H),3.59-3.55 (m, 1H), 3.40-3.15 (m, 3H), 2.17-2.06 (m, 1H), 1.90-1.80 (m,1H), 1.47 (s, 9H).

Step 2

According to a standard Boc-deprotection procedure, pyrrolidine 165 wasprepared in 88% yield.

HCl salt of 172: ¹H NMR (CDCl₃, 400 MHz): δ 11.52 (br s, 1H), 9.23 (brs, 1H), 9.09 (br s, 1H), 8.77 (br s, 1H), 8.54 (d, J=6.4 Hz, 2H),8.25-8.21 (m, 4H), 7.68 (d, J=8.4 Hz, 2H), 7.59 (d, J=8.4 Hz, 2H), 5.36(s, 2H), 5.17 (s, 2H), 4.30-4.25 (m, 1H), 3.97 (d, J=4.8 Hz, 2H),3.33-3.12 (m, 3H), 3.00-2.93 (m, 1H), 2.12-2.03 (m, 1H), 1.83-1.75 (m,1H).

Step 3

General Method A for the Pd coupling reaction afforded product 166 in71% yield.

¹H NMR (CDCl₃, 400 MHz): δ 11.46 (br s, 1H), 8.99 Br s, 1H), 8.20-8.14(m, 6H), 7.63 (d, J=8.8 Hz, 2H), 7.53-7.49 (m, 1H), 5.41 (d, J=14.0 Hz,1H), 5.27 (s, 2H), 5.24 (d, J=14.0 Hz, 1H), 5.17 (s, 2H), 4.42-4.38 (m,1H), 4.27-4.16 (m, 2H), 4.08-3.99 (m, 3H), 3.90-3.80 (m, 2H), 3.56-3.46(m, 1H), 3.34-3.20 (m, 3H), 2.75-2.61 (m, 1H), 2.51-2.43 (m, 1H),2.28-2.20 (m, 1H), 1.22 (d, J=6.0 Hz, 3H), 1.13 (d, J=7.2 Hz, 3H), 0.90(t, J=8.0 Hz, 9H), 0.57 (q, J=8.0 Hz, 6H).

Step 4

General method for the removal of the TES protecting group with 0.06 NHCl (described in step 6 of Scheme 33) gave the OH-product, which wasused for next step—PNB-deprotection (described in general MethodF)—without further isolation or purification to obtain the desired finalproduct 167 in 20% yield.

¹H NMR (D₂O, 400 MHz): δ 4.03-3.94 (m, 2H), 3.89 (dd, J=8.8, 2.8 Hz,1H), 3.83-3.77 (m, 1H), 3.60 (d, J=8.8 Hz, 2H), 3.11 (dd, J=6.4, 2.4 Hz,1H), 3.00-2.98 (m, 1H), 2.78-2.69 (m, 2H), 2.56-2.21 (br m, 3H),2.09-2.04 (m, 1H), 1.76-1.67 (m, 1H), 1.06 (d, J=6.4 Hz, 3H), 0.87 (d,J=7.2 Hz, 3H).

Example 33 Synthesis of Compound 176 & 178

Step 1

To solution of 3-pyrroline (4.68 g, 67.8 mmol) in CH₂Cl₂ (100 mL) at 0°C. was added Et₃N (14.2 mL, 102 mmol), followed by dropwise addition ofa solution of (Boc)₂O (16.28 g, 74.6 mol) in CH₂C₂ (25 mL). Afteraddition, the reaction mixture was stirred at room temperature for 15 h.Then the reaction mixture was treated with H₂O and separated. Afterextraction twice with CH₂Cl₂, the combined organic layers dried overNa₂SO₄, and concentrated in vacuo to give compound 168 (crude 12.2 g).

¹H NMR (CDCl₃, 400 MHz): δ 5.82-5.70 (m, 2H), 4.18-4.03 (m, 4H), 1.46(s, 9H).

Step 2

The Boc-protected pyrroline 168 (crude 12.2 g, 67.8 mmol) was dissolvedin CH₂Cl₂ (200 mL), and m-CPBA (maximum 77% pure, 22.56 g, 101 mmol) wasadded in portions. After the mixture was stirred at room temperature for3 days, the precipitate was filtered off and the filtrate was treatedwith 6N NaOH to PH≈9, after separation, the aqueous phase was extractedtwice with CH₂Cl₂. The combined organic layers washed with brine,concentrated and purified by a silica gel column chromatography withhexane-EtOAc (from 7:3 to 1:1) to afford a pale yellow oil 169 (7.93 g,66% yield over 2 steps).

¹H NMR (CDCl₃, 400 MHz): δ 3.83-3.67 (m, 4H), 3.33-3.30 (m, 2H), 1.45(s, 9H).

Step 3

A solution of the epoxide 169 (6.48 g, 35 mmol) in a mixture of 28%NH₄OH (70 ml) and MeOH (70 ml) in a sealed thick-wall reactor wasallowed to stand at 65° C. for 40 h. Then the mixture was cooled down toroom temperature and the solvent was evaporated. The oily racemicmixture (±)-170 (7.70 g) was used directly for next step without furtherpurification.

¹H NMR (CDCl₃, 400 MHz): δ 4.00 (m, 1H), 3.66 (m, 2H), 3.30 (m, 2H),3.12 (m, 1H), 2.10 (br s, 3H), 1.46 (s, 9H).

Step 4

-   -   To a solution of hydroxylamine (±)-170 (crude 7.70 g, 35 mmol)        in THF (250 ml) was added bis-PNB protected methyl thiourea        guanidine 59 (15.37 g, 31.5 mmol), the reaction mixture was aged        at rt. for 4 days and then concentrated. The residue was        purified by silica gel column chromatography with MeOH—CH₂Cl₂        (from 2:98 to 5:95) to give (±)-171 (15.33 g, 80% yield) as a        white solid.

¹H NMR (CDCl₃, 400 MHz): δ 11.70 (d, J=10.7 Hz, 1H), 8.50 (d, J=14.8 Hz,1H), 8.24 (dd, J=9.0, 8.8 Hz, 4H), 7.54 (dd, J=8.8, 5.9 Hz, 4H), 5.29(s, 2H), 5.21 (s, 2H), 4.32-4.20 (m, 2H), 3.98-3.74 (m, 2H), 3.34-3.20(m, 2H).

Step 5

A 500 ml flask loaded with CH₂Cl₂ (150 mL) was cooled with ice-bath, tothis was added TFA (17.3 ml, 0.23 mol) followed by compound (±)-171(9.03 g, 15 mmol) as a solid. The reaction mixture was aged at 0° C.overnight and then concentrated in vacuo. The concentrate was washedwith hexane and dry Et₂O to afford TFA•salt (±)-172 (8.68 g).

¹H NMR (CDCl₃, 400 MHz): δ 11.50 (s, 1H), 10.20 (br s, 2H), 8.37 (d,J=6.3 Hz, 1H), 8.08 (t, J=8.8 Hz, 4H), 7.42 (dd, J=8.8, 3.0 Hz, 4H),5.15 (s, 2H), 5.08 (s, 2H), 4.42-4.27 (m, 2H), 3.60 (dd, J=12.4, 6.6 Hz,1H), 3.46-3.30 (m, 2H), 3.16 (dd, J=12.4, 2.0 Hz, 1H).

Step 6

According to General Method D, CPI (1.8 g, 3.0 mmol), side chain 172(1.85 g, 3.0 mmol), Pd₂(dba)₃CHCl₃ (199 mg, 0.192 mmol) and P(OEt)₃ (203μL, 1.18 mmol) in THF/Toluene (10/90 mL) were reacted overnight toafford the desired TES product 173 (3S,4S) (less polar isomer: 1.3 g,45%) and 174 (3R,4R) (polar isomer: 1.38 g, 48%).

173 (3S,4S): ¹H NMR (CDCl₃, 400 MHz): δ 11.63 (br s, 1H), 8.42 (d, J=4.4Hz, 1H), 8.22 (m, 6H), 7.66 (d, J=8.4 Hz, 2H), 7.53 (d, J=9.2 Hz, 2H),7.51 (d, J=8.8 Hz, 2H), 5.44 (d, J=14.0 Hz, 1H), 5.28-5.16 (m, 5H), 4.89(s, 1H), 4.25 (t, J=6.0 Hz, 1H), 4.19 (dd, J=6.4, 2.8 Hz, 1H), 4.16-4.02(m, 2H), 3.88 (d, J=14.4 Hz, 1H), 3.32 (d, J=14.4 Hz, 1H), 3.31 (m, 1H),3.24 (dd, J=5.6, 2.8 Hz, 1H), 3.09 (dd, J=9.6, 7.6 Hz, 1H), 2.97 (dd,J=10.0, 7.2 Hz, 1H), 2.62 (dd, J=10.0, 4.8 Hz, 1H), 2.44 (dd, J=9.6, 6.4Hz, 1H), 1.25 (d, J=6.4 Hz, 3H), 1.17 (d, J=7.2 Hz, 3H), 0.94 (t, J=8.0Hz, 9H), 0.60 (q, J=8.0 Hz, 6H).

174 (3R,4R): ¹H NMR (CDCl₃, 400 MHz): δ 11.64 (br s, 1H), 8.42 (d, J=4.4Hz, 1H), 8.22 (m, 6H), 7.66 (d, J=8.8 Hz, 2H), 7.53 (d, J=8.4 Hz, 2H),7.51 (d, J=8.8 Hz, 2H), 5.44 (d, J=13.6 Hz, 1H), 5.33-5.16 (m, 5H), 4.95(s, 1H), 4.25 (t, J=6.0 Hz, 1H), 4.21 (dd, J=6.4, 2.8 Hz, 1H), 4.16-4.02(m, 2H), 3.86 (d, J=14.4 Hz, 1H), 3.32 (d, J=14.0 Hz, 1H), 3.31 (m, 1H),3.24 (dd, J=5.6, 2.8 Hz, 1H), 3.06 (dd, J=9.6, 7.6 Hz, 1H), 2.86 (dd,J=10.0, 7.2 Hz, 1H), 2.70 (dd, J=10.0, 4.4 Hz, 1H), 2.52 (dd, J=9.6, 7.2Hz, 1H), 1.25 (d, J=6.4 Hz, 3H), 1.16 (d, J=7.2 Hz, 3H), 0.94 (t, J=8.0Hz, 9H), 0.60 (q, J=8.0 Hz, 6H).

Step 7

According to General Method E, TES compound 173 (3S,4S) (1.3 g, 1.35mmol), Me₄NF.4H₂O (0.40 g, 2.42 mmol), AcOH (350 μL, 5.83 mmol) in THF(20 mL) and IPA (5 mL) were reacted for 24 h to afford the desired OHproduct 175 (3S,4S) (1.0 g, 85%).

¹H NMR (CDCl₃, 400 MHz): δ 11.62 (br s, 1H), 8.44 (d, J=4.4 Hz, 1H),8.23 (d, J=8.8 Hz, 2H), 8.22 (d, J=8.8 Hz, 2H), 8.19 (d, J=7.6 Hz, 2H),7.64 (d, J=8.8 Hz, 2H), 7.52 (d, J=8.4 Hz, 2H), 7.50 (d, J=8.4 Hz, 2H),5.46 (d, J=14.0 Hz, 1H), 5.27 (s, 2H), 5.20 (d, J=15.2 Hz, 1H), 5.19 (s,2H), 4.89 (s, 1H), 4.24 (t, J=6.4 Hz, 1H), 4.20 (dd, J=10.0, 6.8 Hz,1H), 4.16-4.02 (m, 2H), 3.87 (d, J=14.4 Hz, 1H), 3.35 (m, 1H), 3.31 (d,J=14.4 Hz, 1H), 3.24 (dd, J=6.4, 2.8 Hz, 1H), 3.04 (dd, J=9.6, 7.6 Hz,1H), 2.98 (dd, J=9.6, 7.2 Hz, 1H), 2.54 (dd, J=9.6, 4.8 Hz, 1H), 2.45(dd, J=9.6, 5.6 Hz, 1H), 1.31 (d, J=6.0 Hz, 3H), 1.17 (d, J=7.2 Hz, 3H).

Step 9

According to General Method H, OH compound 175 (3S,4S) (0.44 g, 0.50mmol), 5% Pt/C (300 mg) in IPA (5 mL), THF (12 mL), DI water (4 mL) andpH=6 buffer (6 mL) were reacted for 0.5 h to afford the desired product176 (3S,4S) (60 mg, 31%).

¹H NMR (D₂O, 400 MHz): δ 4.09-3.98 (m, 3H), 3.65 (q, J=5.2 Hz, 1H), 3.55(d, J=13.2 Hz, 1H), 3.21 (dd, J=6.0, 3.2 Hz, 1H), 3.13 (d, J=13.2 Hz,1H), 3.02 (dd, J=9.6, 7.2 Hz, 1H), 2.94 (dd, J=10.8, 8.0 Hz, 1H), 2.86(dd, J=10.8, 7.2 Hz, 1H), 2.39 (m, 2H), 1.08 (d, J=6.4 Hz, 3H), 0.90 (d,J=7.2 Hz, 3H).

Step 10

According to General Method E, TES compound 174 (3R,4R) (1.38 g, 1.44mmol), Me₄NF.4H₂O (0.36 g, 2.18 mmol), AcOH (343 μL, 5.72 mmol) in THF(20 mL) and IPA (5 mL) were reacted for 24 h to afford the desired OHproduct 177 (3R,4R) (1.14 g, 91%).

¹H NMR (CDCl₃, 400 MHz): δ 11.63 (br s, 1H), 8.42 (d, J=4.0 Hz, 1H),8.25-8.17 (m, 6H), 7.64 (dd, J=8.8, 3.6 Hz, 2H), 7.55-7.49 (m, 4H), 5.47(d, J=14.4 Hz, 1H), 5.28 (s, 2H), 5.23-5.19 (m, 3H), 4.92 (s, 1H),4.26-4.20 (m, 2H), 4.16-4.02 (m, 2H), 3.86 (d, J=14.0 Hz, 1H), 3.37 (d,J=13.6 Hz, 1H), 3.28 (m, 1H), 3.26 (m, 1H), 3.04 (m, 2H), 2.87 (dd,J=9.6, 7.2 Hz, 1H), 2.66 (m, 1H), 2.50 (dd, J=9.6, 5.6 Hz, 1H), 1.31 (d,J=6.0 Hz, 3H), 1.17 (d, J=7.2 Hz, 3H).

Step 11

According to General Method H, OH compound 177 (3R,4R) (0.50 g, 0.57mmol), 5% Pt/C (400 mg) in IPA (5 mL), THF (12 mL), DI water (6 mL) andpH=6 buffer (4 mL) were reacted for 0.5 h to afford the desired product176 (3R,4R) (75 mg, 35%).

¹H NMR (D₂O, 400 MHz): δ 4.06-3.96 (m, 3H), 3.65 (br s, 1H), 3.51 (d,J=13.2 Hz, 1H), 3.21 (dd, J=6.0, 2.8 Hz, 1H), 3.16 (d, J=13.2 Hz, 1H),3.00 (dd, J=10.0, 6.8 Hz, 1H), 2.84 (dd, J=10.8, 8.0 Hz, 1H), 2.77 (dd,J=10.8, 7.2 Hz, 1H), 2.40 (m, 2H), 1.07 (d, J=6.0 Hz, 3H), 0.90 (d,J=7.2 Hz, 3H).

Example 34 Synthesis of Compound 185a & 185b

Step 1

To a solution of epoxide 169 (4.0 g, 21.6 mmol) in dry DMSO (20 mL) wasadded KCN (2.8 g, 43.2 mmol), and the reaction mixture was stirred at90° C. for 4 days. After cooling down to room temperature, the mixturewas diluted with ethyl acetate (100 mL) and washed with water (100 mL).The organic phase was separated, washed again with brine, and dried overanhydrous MgSO₄. After concentration, the crude was purified by a flashcolumn chromatography (EtOAc/Hexane 1:1) to afford the desired compound179 (2.0 g, 48%) as colorless oil.

¹H NMR (CDCl₃, 400 MHz) δ, 4.60 (m, 1H), 3.78-3.60 (m, 3H), 3.40 (m,1H), 3.04 (m, 1H), 2.42 (br s, 1H), 1.49 (s, 9H)

Step 2: General Procedures for Hydroxyamilation of Nitrile

A solution of 93 (2.80 g, 14.2 mmol), hydroxylamine hydrochloride (1.98g, 28.4 mmol) and potassium carbonate (3.92 g, 28.4 mmol) in absoluteethanol (50 mL) was heated under reflux for 3 h and allowed to stir atroom temperature overnight and filtered. Evaporation of the filtrate andpurification of the residue by flash chromatography using ethyl acetateas the elute gave 2.50 g (78%) of 180a as foam.

¹H NMR (CDCl₃, 400 MHz) δ 4.51 (s, 2H), 3.59-3.30 (m, 5H), 2.89 (m, 1H),2.11-2.01 (m, 2H), 1.41 (s, 9H)

According to the general procedure as above, 179 (2.0 g, 9.44 mmol),NH₂OH.HCl (2.0 g, 28 mmol) and K₂CO₃ (3.88 g, 28 mmol) were reacted inethanol to afford the desired product 180b (2.0 g, 86%).

¹H NMR (DMSO, 400 MHz) δ 9.00 (br s, 1H), 5.38 (br s, 2H), 5.19 (m, 1H),4.22 (m, 1H), 4.00 (m, 1H), 3.41-3.20 (m, 2H), 3.07 (m, 1H), 2.60 (m,1H), 1.39 (s, 9H)

Step 3: General Procedures for De-Hydroxylation & Protection

A mixture of 180a (2.10 g, 10.5 mmol), acetic anhydride (1.98 mL, 21mmol), Pd/C (5%, 210 mg) and acetic acid (0.5 mL) in ethanol (100 mL)was shaken under hydrogen (50 psi) on a Parr hydrogenator for 15 h. Thecatalyst was removed by filtration and the filtrate was evaporated todryness. The residue was dissolved in a mixture of dry ethanol (50 mL)and toluene (50 mL), and evaporated to dryness. This process wasrepeated three times and the crude amidine residue was used withoutfurther purification.

To a cooled (0° C.) solution of amidine salt and sodium bicarbonate(1.93 g, 23.1 mmol) in dichloromethane/water (30 mL/30 mL) was added4-nitrobenzyl chloroformate (2.71 g, 12.6 mmol). After stirring at roomtemperature for 2 h, the layers were separated and the aqueous wasextracted with CH₂Cl₂ (3×25 mL). The combined organics were dried overNaSO₄ and concentrated to afford crude material. Flash columnchromatography (EtOAc/Hexane 3:1) provided title compound 181a (1.60 g,41%) as white foam solid.

¹H NMR (CDCl₃, 400 MHz) δ 8.85 (br s, 1H), 8.72 (br s, 1H), 8.22 (d,J=7.2 Hz, 2H), 7.60 (d, J=7.2 Hz, 2H), 5.08 (s, 2H), 3.40 (m, 2H), 3.20(m, 2H), 2.99 (m, 1H), 2.03-1.82 (m, 2H), 1.39 (s, 9H).

According to the general procedure as above, 181b was prepared with 31%yield (1 g) from 2.0 g of 180b.

¹H NMR (CDCl₃, 400 MHz) δ 9.26 (br s, 1H), 8.12 (d, J=7.2 Hz, 2H), 7.59(d, J=7.2 Hz, 2H), 5.10 (s, 2H), 4.50 (m, 1H), 3.79 (m, 2H), 3.52 (m,2H), 3.20 (m, 1H), 2.91 (m, 1H), 1.39 (s, 9H).

Step 4

Compound 181a (1.60 g, 4.23 mmol) was added portionwise to a stirred,cooled (0° C.) solution of trifluoroacetic acid (5.0 mL) in CH₂Cl₂ (20mL). This mixture was stirred under nitrogen for 2 h and then evaporatedto dryness. The residue was purified by flash chromatography usingCH₂Cl₂: MeOH (7:3) as the elute to yield trifluoroacetic acid salt 182a(1.5 g, 92%) as foam.

¹H NMR (DMSO, 400 MHz) δ 8.10 (d, J=7.2 Hz, 2H), 7.63 (d, J=7.2 Hz, 2H),5.13 (s, 2H), 3.99 (br s, 4H), 3.40-3.18 (m, 5H), 2.28 (m, 1H), 2.03 (m,1H).

In a similar manner as above, 181b (1.0 g, 2.53 mmol) andtrifluoroacetic acid (2.9 mL, 37.95 mmol) were reacted to afford thedesired product 182b (0.9 g, 88%).

¹H NMR (CDCl₃, 400 MHz) δ 8.10 (d, J=7.2 Hz, 2H), 7.63 (d, J=7.2 Hz,2H), 5.13 (s, 2H), 4.20 (br s, 4H), 3.88-3.52 (m, 4H), 3.00 (m, 2H).

Step 5

According to General Method D, CPI (524 mg, 0.89 mmol), side chain 182a(360 mg, 0.89 mmol), DIEA (154 mL, 0.89 mmol) Pd₂(dba)₃CHCl₃ (83.5 mg,0.081 mmol) and P(OEt)₃ (87 μL, 0.51 mmol) in THF/Toluene (3/25 mL) werereacted for 5 h to afford the desired TES product 183a (0.3 g, 44%).

¹H NMR (CDCl₃, 400 MHz) δ 8.88 (br s, 1H), 8.20 (m, 4H), 7.98 (br s,1H), 7.65 (d, J=8.8 Hz, 2H), 7.56 (d, J=8.4 Hz, 2H), 5.43 (d, J=13.6 Hz,1H), 5.21 (d, J=14.0 Hz, 1H), 5.20 (s, 2H), 4.28-4.22 (m, 2H), 3.95 (d,J=13.6 Hz, 0.7H), 3.90 (d, J=14.4 Hz, 0.3H), 3.42 (d, J=14.0 Hz, 0.3H),3.32 (d, J=14.0 Hz, 0.7H), 3.27-2.89 (m, 5H), 2.55-2.17 (m, 3H), 1.96(m, 1H), 1.24 (d, J=6.0 Hz, 3H), 1.18 (d, J=7.2 Hz, 3H), 0.95-0.91 (m,9H), 0.62-0.56 (m, 6H)

According to General Method D, CPI (1.2 g, 2.0 mmol), side chain 182b(880 mg, 2.09 mmol), DIEA (450 μL, 2.59 mmole), Pd₂(dba)₃CHCl₃ (145.5mg, 0.14 mmol) and P(OEt)₃ (151 μL, 0.88 mmol) in THF/Toluene (10/40 mL)were reacted for 5 h to afford the desired TES product 183b(diastereomer mixture, 0.33 g, 21%).

¹H NMR (CDCl₃, 400 MHz) δ 8.92 (br s, 1H), 8.22 (d, J=8.8 Hz, 2H), 8.19(d, J=9.6 Hz, 2H), 7.92 (br s, 1H), 7.65 (d, J=8.8 Hz, 2H), 7.53 (d,J=8.0 Hz, 2H), 5.43 (d, J=13.2 Hz, 1H), 5.22 (d, J=13.2 Hz, 1H), 5.20(s, 2H), 4.45 (br s, 1H), 4.27-4.20 (m, 2H), 4.00 (d, J=13.2 Hz, 0.5H),3.90 (d, J=14.0 Hz, 0.5H), 3.39 (d, J=14.0 Hz, 0.5H), 3.30-2.89 (m,7.5H), 2.41-2.31 (m, 1H), 1.24 (d, J=6.0 Hz, 3H), 1.18 (d, J=7.2 Hz,3H), 0.93 (t, J=8.0 Hz, 9H), 0.59 (q, J=8.0 Hz, 6H)

Step 6

According to General Method E, TES compound 183a (0.6 g, 0.79 mmol),Me₄NF.4H₂O (0.36 g, 2.18 mmol), AcOH (300 μL, 5.00 mmol) in THF (20 mL)and IPA (5 mL) were reacted for 24 h to afford the desired OH product184a (270 mg, 53%).

¹H NMR (CDCl₃, 400 MHz): δ 8.92 (br s, 1H), 8.20 (m, 4H), 7.98 (br s,1H), 7.66 (d, J=8.8 Hz, 2H), 7.57 (d, J=7.6 Hz, 2H), 5.47 (d, J=13.6 Hz,1H), 5.25-5.17 (m, 3H), 4.30-4.23 (m, 2H), 3.96 (d, J=14.4 Hz, 0.5H),3.91 (d, J=14.8 Hz, 0.5H), 3.43 (d, J=14.8 Hz, 0.5H), 3.34 (d, J=14.0Hz, 0.5H), 3.27-2.89 (m, 6H), 2.55-2.17 (m, 3H), 1.96 (m, 1H), 1.24 (d,J=6.0 Hz, 3H), 1.18 (d, J=7.2 Hz, 3H)

According to General Method E, TES compound 183b (0.33 g, 0.42 mmol),Me₄NF.4H₂O (0.40 g, 2.42 mmol), AcOH (400 μL, 6.67 mmol) in THF (20 mL)and IPA (5 mL) were reacted for 24 h to afford the desired OH product184a (180 mg, 64%).

¹H NMR (CDCl₃, 400 MHz) δ 8.92 (br s, 1H), 8.22-8.17 (m, 4H), 7.92 (brs, 1H), 7.63 (d, J=8.0 Hz, 2H), 7.53 (m, 2H), 5.48-5.43 (m, 1H), 5.22(d, J=14.8 Hz, 1H), 5.19 (s, 2H), 4.42 (br s, 1H), 4.29-4.20 (m, 2H),4.03-3.87 (m, 2H), 3.39-2.72 (m, 8H), 2.41-2.31 (m, 1H), 1.24 (d, J=6.0Hz, 3H), 1.18 (d, J=7.2 Hz, 3H)

Step 7

According to General Method H, OH compound 184a (0.27 g, 0.42 mmol), 5%Pt/C (400 mg) in IPA (5 mL), THF (12 mL), DI water (10 mL) were reactedfor 0.5 h to afford the desired product 185a (35 mg, 25%).

¹H NMR (D₂O, 400 MHz): δ 4.07-3.98 (m, 2H), 3.54 (d, J=14.0 Hz, 1H),3.23 (m, 2H), 3.04 (m, 2H), 2.84-2.53 (m, 4H), 2.14 (br s, 1H), 1.80 (m,1H), 1.08 (d, J=6.4 Hz, 3H), 0.91 (d, J=7.2 Hz, 3H)

According to General Method H, OH compound 184b (0.18 g, 0.27 mmol), 5%Pt/C (400 ing) in IPA (5 mL), THF (12 mL), DT water (10 mL) were reactedfor 0.5 h to afford the desired product 185b (22 mg, 23%).

¹H NMR (D₂O, 400 MHz) δ 4.31 (br s, 1H), 4.03-3.96 (m, 2H), 3.53 (d,J=13.2 Hz, 1H), 3.21 (m, 1H), 3.15 (d, J=13.6 Hz, 1H), 3.03-2.80 (m,5H), 2.64 (m, 1H), 2.39-2.27 (m, 1H), 1.07 (d, J=4.8 Hz, 3H), 0.96-0.88(m, 3H)

Example 35 Dilution Antimicrobial Susceptibility Tests

The MIC (minimum inhibitory concentration) was determined by the NCCLS(National Committee for Clinical Laboratory Standards) methods 2000.Methods for dilution antimicrobial susceptibility testing for bacteriathat growth aerobically (M7-A5, vol. 20, No. 2). The agar dilutionmethod for determining antimicrobial susceptibility was carried outusing Mueller-Hinton agar. A final inoculum of 10⁴ CFU/spot was appliedwith an inoculation device. Broth dilution tests were performed with5×10⁵ CFU/well in 96 well plates. The susceptibilities of streptococciwere determined by Mueller-Hinton agar supplemented with 5% sheep blood.All assays were run with the indicated control strains, available fromthe ATCC (American Type Culture Collection, Rockville, Md.). Results ofthe antimicrobial susceptibility tests of certain compounds againstGram-negative organisms are shown in Table 1.

Abbreviations used in Table 1 are defined as follows: Cf-R: CeftazidimeResistance, Ci-R: Ciprofloxacin Resistance, Gen-R: GentamycinResistance, Imp-R: Imipenem Resistance, Mp-R: Meropenem Resistance,Ofx-R: Ofloxacin Resistance, B+: β-lactamase Production, AmpC:AmpC-lactamase Hyper production, CBPase: Carbapenemase Production.

TABLE 1 Antibacterial activity (MIC^(a)) data on G(−) bacterial strains# Genus Species Resistance ^(b) 7 12 19 27 32 37 43 46 Meropenem 1Acinetobacter calcoaceticus 4 0.5 0.5 0.25 2 2 0.5 1 0.5 2 Acinetobacterbaumannii Cf-R, Ci-R 2 0.5 0.5 0.5 2 4 1 2 1 3 Acinetobacter baumanniiCf-R, Ci-R, 4 2 2 2 4 8 4 4 8 Mp-I 4 Acinetobacter baumannii Cf-R,Imp-I, 4 1 1 1 2 4 4 4 4 CBPase 5 Citrobacter diversus 0.063 0.13 0.250.25 1 0.5 0.25 0.25 0.016 6 Citrobacter freundii 0.13 0.25 0.25 0.25 10.5 0.25 0.5 0.016 7 Enterobacter aerogenes 0.5 1 1 1 4 2 1 1 0.063 8Enterobacter cloacae Cf-R, B+ 0.13 0.13 0.25 0.25 1 0.5 0.25 0.25 0.0169 Enterobacter cloacae Cf-R 0.25 0.25 0.5 0.5 2 1 0.5 0.5 0.063 10Escherichia coli Ci-R 0.13 0.13 0.25 0.25 1 0.5 0.25 0.25 0.016 11Escherichia coli Ci-R, AmpC 0.13 0.13 0.25 0.25 0.5 0.5 0.25 0.25 0.01612 Escherichia coli 0.13 0.13 0.25 0.25 1 0.5 0.25 0.25 0.016 13Escherichia coli 0.13 0.25 0.25 0.25 1 0.5 0.25 0.25 0.016 14 Klebsiellaoxytoca 0.25 0.5 0.5 0.5 2 0.5 0.5 0.5 0.031 15 Klebsiella pneumoniae0.25 0.5 0.5 0.5 2 1 0.5 0.5 0.031 16 Moraxella catarrhalis ≦0.008≦0.008 ≦0.008 ≦0.008 0.016 ≦0.008 ≦0.008 ≦0.008 ≦0.008 17 MorganellaMorganii 1 2 4 4 8 2 2 2 0.13 18 Morganella Morganii 1 1 2 2 4 2 2 20.13 19 Proteus vulgaris 0.5 0.5 1 1 4 1 1 1 0.13 20 Proteus mirabilis0.13 0.25 0.25 0.25 1 0.5 0.25 0.25 0.031 21 Providencia rettgeri 0.5 11 1 4 1 1 1 0.031 22 Salmonella typhimurium 0.13 0.25 0.25 0.25 1 0.50.5 0.25 0.016 23 Serratia marcescens 0.25 0.5 1 0.5 2 1 1 1 0.031 24Serratia marcescens 0.5 1 1 1 2 2 1 1 0.031 25 Shigella dysenteriae 0.250.25 0.25 0.5 1 0.5 0.5 0.5 0.016 26 Shigella sonnei 0.13 0.25 0.25 0.251 0.5 0.25 0.25 0.016 27 Shigella flexneri 0.13 0.25 0.25 0.13 0.5 0.250.25 0.25 0.016 28 Stenotrophomonas maltophilia 0.5 0.25 0.25 0.13 1 10.5 0.5 0.25 29 Pseudomonas aeruginosa 16 4 4 2 16 8 4 4 0.25 30Pseudomonas aeruginosa 8 4 4 2 16 8 2 4 0.5 31 Pseudomonas aeruginosaOfx-R 2 1 1 0.5 4 4 1 1 0.5 32 Pseudomonas aeruginosa Cf-R 8 4 4 2 16 84 4 0.13 33 Pseudomonas aeruginosa Ci-R 8 4 4 2 16 8 4 4 1 34Pseudomonas aeruginosa Gen-R 8 4 4 2 16 8 4 4 0.5 35 Pseudomonasaeruginosa Ip-R 32 8 8 4 32 16 8 16 4 36 Pseudomonas aeruginosa Cf-R,Ci-R, >32 32 32 8 >32 32 16 32 8 Mp-R # Genus Species Resistance 49 5864 67 72 78 83 84 Meropenem 1 Acinetobacter calcoaceticus 0.25 0.5 0.132 1 2 2 0.25 0.5 2 Acinetobacter baumannii Cf-R, Ci-R 0.5 2 1 8 2 2 40.5 1 3 Acinetobacter baumannii Cf-R, Ci-R, 2 4 1 8 4 4 8 1 8 Mp-I 4Acinetobacter baumannii Cf-R, Imp-I, 1 2 0.5 4 4 4 8 1 4 CBPase 5Citrobacter diversus 0.063 0.13 0.5 0.13 0.5 0.5 0.5 0.25 0.016 6Citrobacter freundii 0.13 0.25 0.5 0.25 0.5 0.5 1 0.5 0.016 7Enterobacter aerogenes 0.25 0.5 1 4 1 2 4 1 0.063 8 Enterobacter cloacaeCf-R, B+ 0.063 0.13 0.5 0.13 0.25 0.5 0.5 0.25 0.016 9 Enterobactercloacae Cf-R 0.13 0.25 0.5 0.25 1 1 1 0.5 0.063 10 Escherichia coli Ci-R0.063 0.13 0.25 0.13 0.25 0.5 0.5 0.25 0.016 11 Escherichia coli Ci-R,AmpC 0.063 0.13 0.25 0.13 0.25 0.5 0.5 0.25 0.016 12 Escherichia coli0.063 0.13 0.5 0.13 0.25 0.5 0.5 0.25 0.016 13 Escherichia coli 0.130.25 0.5 0.25 0.5 1 1 0.5 0.016 14 Klebsiella oxytoca 0.25 0.25 1 0.25 11 2 0.5 0.031 15 Klebsiella pneumoniae 0.25 0.25 1 4 0.5 1 1 0.5 0.03116 Moraxella catarrhalis ≦0.008 ≦0.008 ≦0.008 0.016 0.016 0.016 0.031≦0.008 ≦0.008 17 Morganella Morganii 2 1 4 16 4 8 8 4 0.13 18 MorganellaMorganii 1 1 2 16 4 4 8 2 0.13 19 Proteus vulgaris 0.5 0.5 1 16 2 4 4 10.13 20 Proteus mirabilis 0.13 0.25 0.5 0.25 0.5 1 1 0.5 0.031 21Providencia rettgeri 0.5 0.5 2 1 2 4 4 2 0.031 22 Salmonella typhimurium0.13 0.25 0.5 0.25 0.5 1 1 0.5 0.016 23 Serratia marcescens 0.25 0.5 10.5 1 2 2 1 0.031 24 Serratia marcescens 0.25 0.5 2 0.5 1 2 2 1 0.031 25Shigella dysenteriae 0.13 0.25 0.5 0.25 0.5 1 1 0.5 0.016 26 Shigellasonnei 0.13 0.25 0.5 0.25 0.5 1 1 0.5 0.016 27 Shigella flexneri 0.0630.13 0.25 0.13 0.25 0.5 0.5 0.25 0.016 28 Stenotrophomonas maltophilia0.13 0.25 0.25 2 0.5 1 1 0.25 0.25 29 Pseudomonas aeruginosa 4 8 1 1 8 816 2 0.25 30 Pseudomonas aeruginosa 4 8 1 2 8 8 16 2 0.5 31 Pseudomonasaeruginosa Ofx-R 2 4 0.25 2 2 2 8 0.5 0.5 32 Pseudomonas aeruginosa Cf-R4 8 2 2 4 16 16 2 0.13 33 Pseudomonas aeruginosa Ci-R 4 8 2 2 8 16 16 21 34 Pseudomonas aeruginosa Gen-R 4 8 2 2 8 16 32 2 0.5 35 Pseudomonasaeruginosa Ip-R 8 16 2 4 8 16 32 4 4 36 Pseudomonas aeruginosa Cf-R,Ci-R, 32 32 16 16 32 >32 >32 16 8 Mp-R # Genus Species Resistance 89 9399 106 111 115 120 127 Meropenem 1 Acinetobacter calcoaceticus 0.13 10.25 4 1 0.5 4 1 0.5 2 Acinetobacter baumannii Cf-R, Ci-R 1 2 1 2 4 1 81 1 3 Acinetobacter baumannii Cf-R, Ci-R, 1 8 2 8 4 2 16 4 8 Mp-I 4Acinetobacter baumannii Cf-R, Imp-I, 0.5 2 2 2 4 1 4 2 4 CBPase 5Citrobacter diversus 0.13 0.25 0.5 1 0.5 0.25 0.5 0.5 0.016 6Citrobacter freundii 0.25 0.5 1 1 0.5 0.25 0.5 0.5 0.016 7 Enterobacteraerogenes 1 2 2 2 2 1 2 2 0.063 8 Enterobacter cloacae Cf-R, B+ 0.130.25 0.5 1 1 0.13 0.5 0.5 0.016 9 Enterobacter cloacae Cf-R 0.25 0.5 1 21 0.25 1 1 0.063 10 Escherichia coli Ci-R 0.13 0.25 0.5 1 0.5 0.25 0.50.5 0.016 11 Escherichia coli Ci-R, AmpC 0.13 0.25 0.5 0.5 0.5 0.25 0.50.5 0.016 12 Escherichia coli 0.13 0.25 0.5 1 0.5 0.25 0.5 0.5 0.016 13Escherichia coli 0.25 0.5 1 1 0.5 0.25 0.5 0.5 0.016 14 Klebsiellaoxytoca 0.5 0.5 2 2 1 0.25 1 1 0.031 15 Klebsiella pneumoniae 0.25 0.5 11 1 0.5 1 1 0.031 16 Moraxella catarrhalis ≦0.008 0.031 ≦0.008 ≦0.008≦0.008 ≦0.008 0.016 ≦0.008 ≦0.008 17 Morganella Morganii 2 8 4 4 4 2 8 40.13 18 Morganella Morganii 2 4 4 4 2 2 4 4 0.13 19 Proteus vulgaris 0.52 2 2 2 1 4 2 0.13 20 Proteus mirabilis 0.25 0.5 1 1 0.5 0.25 0.5 0.50.031 21 Providencia rettgeri 1 2 4 4 2 1 2 2 0.031 22 Salmonellatyphimurium 0.25 0.5 1 1 0.5 0.25 0.5 0.5 0.016 23 Serratia marcescens0.5 1 2 2 1 0.5 1 1 0.031 24 Serratia marcescens 0.5 1 4 2 2 1 2 2 0.03125 Shigella dysenteriae 0.25 0.5 1 1 0.5 0.25 0.5 1 0.016 26 Shigellasonnei 0.25 0.5 1 1 0.5 0.25 0.5 0.5 0.016 27 Shigella flexneri 0.130.25 1 0.5 0.5 0.25 0.5 0.5 0.016 28 Stenotrophomonas maltophilia 0.130.5 0.25 1 0.5 0.25 1 0.5 0.25 29 Pseudomonas aeruginosa 1 4 2 8 4 4 168 0.25 30 Pseudomonas aeruginosa 1 4 2 8 4 4 16 8 0.5 31 Pseudomonasaeruginosa Ofx-R 0.5 4 1 4 1 1 4 2 0.5 32 Pseudomonas aeruginosa Cf-R 14 4 8 4 4 16 8 0.13 33 Pseudomonas aeruginosa Ci-R 1 8 4 8 8 8 16 8 1 34Pseudomonas aeruginosa Gen-R 1 8 4 8 4 8 16 8 0.5 35 Pseudomonasaeruginosa Ip-R 4 16 4 16 8 8 >32 16 4 36 Pseudomonas aeruginosa Cf-R,Ci-R, 8 16 >32 >32 >32 >32 >32 >32 8 Mp-R # Genus Species Resistance 132137 143 146 151 156 163 167 Meropenem 1 Acinetobacter calcoaceticus 4 18 1 0.25 0.5 2 32 0.5 2 Acinetobacter baumannii Cf-R, Ci-R 16 2 32 1 0.52 4 >32 1 3 Acinetobacter baumannii Cf-R, Ci-R, 16 8 32 2 2 4 8 >32 8Mp-I 4 Acinetobacter baumannii Cf-R, Imp-I, 32 2 >32 4 1 2 16 >32 4CBPase 5 Citrobacter diversus 0.25 0.5 0.5 0.5 0.25 0.5 0.5 1 0.016 6Citrobacter freundii 0.25 1 1 0.5 0.5 0.5 1 2 0.016 7 Enterobacteraerogenes 1 2 4 2 2 2 2 4 0.063 8 Enterobacter cloacae Cf-R, B+ 0.25 0.50.5 0.25 0.25 0.5 0.5 2 0.016 9 Enterobacter cloacae Cf-R 0.5 1 1 1 0.51 1 2 0.063 10 Escherichia coli Ci-R 0.25 0.5 0.5 0.25 0.25 0.5 0.5 10.016 11 Escherichia coli Ci-R, AmpC 0.25 0.5 0.5 0.25 0.25 0.5 0.5 10.016 12 Escherichia coli 0.25 0.5 0.5 0.5 0.25 0.5 0.5 1 0.016 13Escherichia coli 0.25 1 0.5 0.5 0.5 0.5 0.5 1 0.016 14 Klebsiellaoxytoca 0.5 1 1 1 0.5 1 1 2 0.031 15 Klebsiella pneumoniae 0.5 1 2 1 0.51 1 2 0.031 16 Moraxella catarrhalis 0.016 ≦0.008 0.063 0.016 ≦0.008≦0.008 ≦0.008 2 ≦0.008 17 Morganella Morganii 4 4 16 8 4 4 8 8 0.13 18Morganella Morganii 4 4 8 8 4 4 8 8 0.13 19 Proteus vulgaris 1 2 8 4 1 24 4 0.13 20 Proteus mirabilis 0.25 0.5 1 0.5 0.5 0.5 0.5 2 0.031 21Providencia rettgeri 1 2 2 2 2 2 1 2 0.031 22 Salmonella typhimurium0.25 1 0.5 0.5 0.5 1 0.5 2 0.016 23 Serratia marcescens 0.5 2 1 2 1 2 22 0.031 24 Serratia marcescens 1 2 2 2 1 2 2 4 0.031 25 Shigelladysenteriae 0.25 1 1 0.5 0.5 1 1 1 0.016 26 Shigella sonnei 0.25 1 1 0.50.5 0.5 0.5 1 0.016 27 Shigella flexneri 0.13 0.5 0.5 0.25 0.25 0.5 0.51 0.016 28 Stenotrophomonas maltophilia 2 0.5 4 0.5 0.25 0.5 1 16 0.2529 Pseudomonas aeruginosa >32 8 >32 8 2 4 16 >32 0.25 30 Pseudomonasaeruginosa >32 8 >32 8 2 4 16 >32 0.5 31 Pseudomonas aeruginosa Ofx-R 324 32 2 1 2 4 32 0.5 32 Pseudomonas aeruginosa Cf-R >32 8 >32 8 2 816 >32 0.13 33 Pseudomonas aeruginosa Ci-R >32 16 >32 8 2 8 16 >32 1 34Pseudomonas aeruginosa Gen-R >32 16 >32 8 2 8 16 >32 0.5 35 Pseudomonasaeruginosa Ip-R >32 32 >32 16 4 8 16 >32 4 36 Pseudomonas aeruginosaCf-R, Ci-R, >32 >32 >32 32 8 >32 >32 >32 8 Mp-R # Genus SpeciesResistance 176 178 185a 185b Meropenem 1 Acinetobacter calcoaceticus0.13 0.13 0.13 0.13 0.5 2 Acinetobacter baumannii Cf-R, Ci-R 0.5 1 0.50.50 1 3 Acinetobacter baumannii Cf-R, Ci-R, 1 1 1 2 8 Mp-I 4Acinetobacter baumannii Cf-R, Imp-I, 1 1 1 1 4 CBPase 5 Citrobacterdiversus 0.063 0.25 0.13 0.063 0.016 6 Citrobacter freundii 0.13 0.50.13 0.063 0.016 7 Enterobacter aerogenes 0.25 1 0.5 0.25 0.063 8Enterobacter cloacae Cf-R, B+ 0.063 0.25 0.063 0.063 0.016 9Enterobacter cloacae Cf-R 0.13 0.5 0.13 0.13 0.063 10 Escherichia coliCi-R 0.063 0.25 0.063 0.031 0.016 11 Escherichia coli Ci-R, AmpC 0.0630.25 0.063 0.031 0.016 12 Escherichia coli 0.063 0.5 0.063 0.031 0.01613 Escherichia coli 0.13 0.5 0.13 0.063 0.016 14 Klebsiella oxytoca 0.130.5 0.25 0.13 0.031 15 Klebsiella pneumoniae 0.13 0.5 0.25 0.13 0.031 16Moraxella catarrhalis ≦0.008 ≦0.008 ≦0.008 ≦0.008 ≦0.008 17 MorganellaMorganii 1 2 1 0.5 0.13 18 Morganella Morganii 0.5 2 1 0.5 0.13 19Proteus vulgaris 0.25 2 0.5 0.25 0.13 20 Proteus mirabilis 0.063 0.50.13 0.063 0.031 21 Providencia rettgeri 0.25 1 0.5 0.25 0.031 22Salmonella typhimurium 0.13 0.5 0.13 0.063 0.016 23 Serratia marcescens0.25 1 0.25 0.13 0.031 24 Serratia marcescens 0.25 2 0.5 0.13 0.031 25Shigella dysenteriae 0.13 0.5 0.13 0.063 0.016 26 Shigella sonnei 0.0630.5 0.13 0.063 0.016 27 Shigella flexneri 0.063 0.25 0.063 0.063 0.01628 Stenotrophomonas maltophilia 0.063 0.13 0.063 0.13 0.25 29Pseudomonas aeruginosa 1 1 1 1 0.25 30 Pseudomonas aeruginosa 1 2 1 10.5 31 Pseudomonas aeruginosa Ofx-R 0.5 1 0.5 0.5 0.5 32 Pseudomonasaeruginosa Cf-R 1 2 1 1 0.13 33 Pseudomonas aeruginosa Ci-R 1 2 1 1 1 34Pseudomonas aeruginosa Gen-R 1 2 1 1 0.5 35 Pseudomonas aeruginosa Ip-R2 2 2 4 4 36 Pseudomonas aeruginosa Cf-R, Ci-R, 8 16 8 8 8 Mp-R

The compositions, methods and/or processes disclosed and claimed hereincan be made and executed without undue experimentation in light of thepresent disclosure. While the compositions and methods of this inventionhave been described in terms of preferred embodiments, it will beapparent to those of skill in the art that variations may be applied tothe compositions, methods and/or processes and in the steps or in thesequence of steps of the methods described herein without departing fromthe concept and scope of the invention. More specifically, it will beapparent that certain agents which are both chemically andphysiologically related may be substituted for the agents describedherein while the same or similar results would be achieved. All suchsimilar substitutes and modifications apparent to those skilled in theart are deemed to be within the scope and concept of the invention.

1-21. (canceled)
 22. A method of treating a bacterial infection in ahost comprising administering a therapeutic amount of a compound ofFormula I:

or a pharmaceutically acceptable salt, ester or prodrug thereof, whereinR¹ and R² are each independently selected from H or alkyl; P is H, OH,halogen, or hydroxyl protected by a hydroxyl protecting group; n is 0, 1or 2; X is —(CR₂)_(m)— or —C(═O)—; m is 0, 1 or 2; Y is CN, OR, SR′ orNRR′; each R is independently selected from H, alkyl or haloalkyl; R′ isH, alkyl, NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂; CR₂C(═O)NR₂;C(═NR)R; C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; orC(═O)CR₂NRC(═NR)NR₂; and Z is H, alkyl, halo, CN, OR, SR′ or NRR′;optionally in a pharmaceutically acceptable carrier or diluent.
 23. Amethod of treating a bacterial infection in a host comprisingadministering a therapeutic amount of a compound of Formula II:

or a pharmaceutically acceptable salt, ester or prodrug thereof, whereinR¹ and R² are each independently selected from H or alkyl; P is H, OH,halogen, or hydroxyl protected by a hydroxyl protecting group; X is—(CR₂)_(m)— or —C(═O)—; m is 0, 1 or 2; Y is CN, OR, SR′ or NRR′; each Ris independently selected from H, alkyl or haloalkyl; R′ is H, alkyl,NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂; CR₂C(═O)NR₂; C(═NR)R;C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; or C(═O)CR₂NRC(═NR)NR₂;and Z is H, alkyl, halo, CN, OR, SR′ or NRR′; optionally in apharmaceutically acceptable carrier or diluent.
 24. A method of treatinga bacterial infection in a host comprising administering a therapeuticamount of a compound of Formula III:

or a pharmaceutically acceptable salt, ester or prodrug thereof, whereinR¹ and R² are each independently selected from H or alkyl; P is H, OH,halogen, or hydroxyl protected by a hydroxyl protecting group; X is—(CR₂)_(m)— or —C(═O)—; m is 0, 1 or 2; Y is CN, OR, SR′ or NRR′; each Ris independently selected from H, alkyl or haloalkyl; R′ is H, alkyl,NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂; CR₂C(═O)NR₂; C(═NR)R;C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; or C(═O)CR₂NRC(═NR)NR₂;and Z is H, alkyl, halo, CN, OR, SR′ or NRR′; optionally in apharmaceutically acceptable carrier or diluent.
 25. A method of treatinga bacterial infection in a host comprising administering a therapeuticamount of a compound of Formula IV:

or a pharmaceutically acceptable salt, ester or prodrug thereof, whereinR¹, R² and R³ are each independently selected from H or alkyl; P is H,OH, halogen, or hydroxyl protected by a hydroxyl protecting group; n is0, 1, or 2 X is —(CR₂)_(m)— or —C(═O)—; m is 0, 1 or 2; Y is CN, OR, SR′or NRR′; each R is independently selected from H, alkyl or haloalkyl; R′is H, alkyl, NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂;CR₂C(═O)NR₂; C(═NR)R; C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; orC(═O)CR₂NRC(═NR)NR₂; and Z is H, alkyl, halo, CN, OR, SR′ or NRR′;optionally in a pharmaceutically acceptable carrier or diluent.
 26. Amethod of treating a bacterial infection in a host comprisingadministering a therapeutic amount of a compound of Formula V:

or a pharmaceutically acceptable salt, ester or prodrug thereof, whereinR¹, R² and R³ are each independently selected from H or alkyl; P is H,OH, halogen, or hydroxyl protected by a hydroxyl protecting group; X is—(CR₂)_(m)— or —C(═O)—; m is 0, 1 or 2; Y is CN, OR, SR′ or NRR′; each Ris independently selected from H, alkyl or haloalkyl; R′ is H, alkyl,NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂; CR₂C(═O)NR₂; C(═NR)R;C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; or C(═O)CR₂NRC(═NR)NR₂;and Z is H, alkyl, halo, CN, OR, SR′ or NRR′; optionally in apharmaceutically acceptable carrier or diluent.
 27. A method of treatinga bacterial infection in a host comprising administering a therapeuticamount of a compound of Formula VI:

or a pharmaceutically acceptable salt, ester or prodrug thereof, whereinR¹ and R² are each independently selected from H or alkyl; P is H, OH,halogen, or hydroxyl protected by a hydroxyl protecting group; X is—(CR₂)_(m)— or —C(═O)—; m is 0, 1 or 2; Y is CN, OR, SR′ or NRR′; each Ris independently selected from H, alkyl or haloalkyl; R′ is H, alkyl,NR₂; C(═O)R; SO₂R; SO₂NR₂; C(═NR)NR₂; C(═O)NR₂; CR₂C(═O)NR₂; C(═NR)R;C(═NR)NRSO₂R; C(═NR)NRC(═O)R; C(═O)CR₂NRSO₂NR₂; or C(═O)CR₂NRC(═NR)NR₂;and Z is H, alkyl, halo, CN, OR, SR′ or NRR′.
 28. A method of treating abacterial infection in a host comprising administering a therapeuticamount of a compound selected from the group consisting of:


29. A method of treating a bacterial infection in a host comprisingadministering a therapeutic amount of a compound selected from the groupconsisting of:


30. The method of claim 22, wherein the host is a human.
 31. The methodof claim 22, wherein the compound is administered orally, parenterally,intravenously, intradermally, subcutaneously or topically.
 32. Themethod of claim 22, wherein the bacterial infection is due togram-negative bacteria.
 33. The method of claim 22, wherein thebacterial infection is a drug resistant or multiple-drug resistantbacterial infection.
 34. The method of claim 22, where in the compoundis administered in combination or alternation with anotheranti-bacterial agent.
 35. The method of claim 34, wherein the otheranti-bacterial agent is a β-lactamase inhibitor.